LOCUS       HMBP-3C-STAR.66-285       7339 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses MBP-tagged STARD1 protein domain (residues 66-285).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7339)
  AUTHORS   Sluchanko NN, Tugaeva KV, Faletrov YV, Levitsky DI
  TITLE     High-yield soluble expression, purification and characterization of 
            human steroidogenic acute regulatory protein (StAR) fused to a 
            cleavable Maltose-Binding Protein (MBP).
  JOURNAL   Protein Expr Purif. 2016 Mar;119:27-35. doi: 
            10.1016/j.pep.2015.11.002. Epub 2015 Nov 7.
  PUBMED    26555181
REFERENCE   2  (bases 1 to 7339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr Purif."; date: "2016-03"; volume: "119:27-35. doi"; pages: " 
            10.1016/j.pep.2015.11.002. Epub 2015 Nov 7"
COMMENT     Sequence Label: HMBP-3C-STAR.66-285
FEATURES             Location/Qualifiers
     source          1..7339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             126..143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             144..1244
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1101..1110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1218..1241
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1293..1304
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1317..1340
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2025..2042)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2025..2041)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2034..2056)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2220..2237)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2390..2476
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2568..2595
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2614..2705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2706..2774,2775..3566)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2924..2943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3608..4121)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3803..3824)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4015..4034
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4232..4820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4721..4740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4974..4991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5006..5148
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5074..5093)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5234..5256
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5250..5441)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        5937..6014
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6015..7097
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(6034..6053)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccatagc
      121 atatgcatca ccatcaccat cacatgaaaa ctgaagaagg taaactggta atctggatta
      181 acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg
      241 gaattaaagt caccgttgag catccggata aactggaaga gaaattccca caggttgcgg
      301 caactggcga tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc
      361 aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt
      421 ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag
      481 cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga
      541 tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc
      601 aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg
      661 aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc
      721 tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca
      781 tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat
      841 ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg
      901 gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga
      961 acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag
     1021 cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg
     1081 cgaaagatcc acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa atcatgccga
     1141 acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca
     1201 gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg agctcgaaca
     1261 acaacaacaa taacaataac aacaacctcg ggatcgaggg aaggatttca gaattcctgg
     1321 aagttctgtt ccaggggccc ggatccactc tctacagtga ccaggagctg gcctatctcc
     1381 agcaggggga ggaggccatg cagaaggcct tgggcatcct tagcaaccaa gagggctgga
     1441 agaaggagag tcagcaggac aatggggaca aagtgatgag taaagtggtc ccagatgtgg
     1501 gcaaggtgtt ccggctggag gtcgtggtgg accagcccat ggagaggctc tatgaagagc
     1561 tcgtggagcg catggaagca atgggggagt ggaaccccaa tgtcaaggag atcaaggtcc
     1621 tgcagaagat cggaaaagat acattcatta ctcacgagct ggctgccgag gcagcaggaa
     1681 acctggtggg gccccgtgac tttgtgagcg tgcgctgtgc caagcgccga ggctccacct
     1741 gtgtgctggc tggcatggcc acagacttcg ggaacatgcc tgagcagaag ggtgtcatca
     1801 gggcggagca cggtcccact tgcatggtgc ttcacccgtt ggctggaagt ccctctaaga
     1861 ccaaacttac gtggctactc agcatcgacc tcaaggggtg gctgcccaag agcatcatca
     1921 accaggtcct gtcccagacc caggtggatt ttgccaacca cctgcgcaag cgcctggagt
     1981 cccaccctgc ctctgaagcc aggtgttgaa gaccaaagct tggcactggc cgtcgtttta
     2041 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2101 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2161 cgcagcctga atggcgaatg gcagcttggc tgttttggcg gatgagataa gattttcagc
     2221 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     2281 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     2341 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     2401 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2461 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     2521 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     2581 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     2641 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2701 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2761 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2821 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2881 gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2941 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3001 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3061 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3121 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3181 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3241 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3301 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3361 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3421 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3481 tctacacgac ggggagtcag gcaactatgg atgaacggaa tagacagatc gctgagatag
     3541 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3601 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     3661 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3721 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3781 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3841 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3901 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3961 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4021 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4081 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     4141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4921 ttgccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4981 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     5041 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5101 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     5161 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5221 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5281 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc
     5341 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     5401 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     5461 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     5521 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     5581 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     5641 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     5701 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     5761 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     5821 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     5881 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga aattccgaca
     5941 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     6001 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     6061 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     6121 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     6181 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     6241 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     6301 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     6361 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     6421 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     6481 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     6541 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     6601 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     6661 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     6721 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     6781 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     6841 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     6901 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     6961 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     7021 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     7081 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcacaa
     7141 ttctcatgtt tgacagctta tcatcgactg cacggtgcac caatgcttct ggcgtcaggc
     7201 agccatcgga agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc
     7261 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
     7321 aatattctga aatgagctg
//