LOCUS       pDawn-sfGFP                7913 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses superfolder GFP under the control of a blue-light 
            transcriptional regulator.
ACCESSION   .
VERSION     .
KEYWORDS    pDawn-sfGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7913)
  AUTHORS   Jin X, Riedel-Kruse IH
  TITLE     Biofilm Lithography enables high-resolution cell patterning via 
            optogenetic adhesin expression.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Apr 3;115(14):3698-3703. doi: 
            10.1073/pnas.1720676115. Epub 2018 Mar 19.
  PUBMED    29555779
REFERENCE   2  (bases 1 to 7913)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..7913
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(1605..1682)
                     /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."
     primer_bind     1888..1907
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             2230..2241
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2248..2958
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             2959..2991
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      3039..3110
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3126..3153
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3229..3251
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3270..3287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3297..3314
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3318..3350
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             3358..3369
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3376..4089
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             4102..4119
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4172..4190)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4186..4233
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4270..4725
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4357..4376)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4567..4588
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4817..5632)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5540..5559
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5624..5643
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5754..6342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6243..6262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6496..6513
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6528..6667
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6593..6612)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6753..6775
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6769..6960)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gatcagccct tccatgacct ggcgctccct ggggctcaac gaggcgacgc gggcggcgat
       61 atcctgcgcg acggcctcgc tcttggcggc cggctcggcc tggcggatcg ccgattcgat
      121 catggcggtg aggcggtcgt cctcgaaagg cttttccaga aagtccaccg cccctaactt
      181 catcgcctcg accgcgagcg gcacgtcgcc gtgaccggtc atgatgagga tcggaaaggg
      241 gctttgctgc gccttcatcc gcttcaacag ctcgatgccg tcaaggcccg gcatgcgcac
      301 gtcggagacg acgcagccga aggagagacc cggcagggcg tcgagaaagg cttgcgcgtc
      361 gtcaaacagc gtgacgccga agccggcaga atccagcagg aaattcagcg aatcccgcat
      421 cgccgcgtcg tcgtcgatga cgtagatatg tcccttggtc gtcatgctag acctcctatc
      481 atctcgtcgg ctgccgggag ggtgaagcgg aaggtcgccc cgcccgatgc gttgctctcg
      541 gcccacatgc gcccgccgtg agcttcgatg atcgagcggc tgatggacag tcccacgccc
      601 atgccggtgt ccttggtggt gaagaaagtc tgaaacaggt tcggaatgac gtcgtcctgg
      661 aaaccgctgc cggtgtcgga cacttcgacc tcgatcatgt cgtcggcggc gggggtgttg
      721 gtgacgacga gctcgcgtcg ctgcgactga gccatcgctt ccagcgcgtt gcggaacagg
      781 ttgaccagga cctgctggat ctgcacccgg tcggcgagaa cgagatcggc gcccggatcg
      841 agactgaagc ggagctgcac gttctgctcg cgcgcgccgg caagcccgag cgcgccggcc
      901 tcctcgatca gcttggagag actctcgacc cgcttctccg attcgccgcg ggcaacgaag
      961 tcgcgcaggc gccggatgat ctggccggcg cgcagcgcct gctcggcggc gcggtccagg
     1021 gcgctttcga ccttcggtgt gttcggatca ctgctgccgg caagcagccg ccgcgagccc
     1081 ttcatgtagt tgctgatcgc cgccagcggc tggttgagct cgtgcgcgag cgcggacgcc
     1141 atttcgccca tggcgctcag cctggagacg tggacgagct cggattgcag ttcctggagg
     1201 cgcgcctggg tctgctggtg ctcggtgata tcattctgaa ttccgacaaa atacgtttta
     1261 tcctctattt ccattggatc aatatttaat tcattccaga acatcgttcc gtcttttttg
     1321 tagttttgga tctgaacggt gaccggttct ttattttgta aagcggttct gatgttgtcc
     1381 acttctgcag gatctgtgtg tttcccctgt aagaagcgac agttctttcc taaaatttcc
     1441 tcggtctcgt agccggtcat ttgaacaaag ccttgattta cgtagacaat aggattatct
     1501 tcaagtgcgg gatctgtaat taccacaccg actcgcacgt gatcaagtgc ttttttgatg
     1561 acttccagct gtcctggtat cccaaatgat tgaaaactag ccacattcac caccctcaat
     1621 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg
     1681 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     1741 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     1801 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     1861 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     1921 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctacgc
     1981 ccgtgatcct gatcaccggc tatccggacg aaaacatctc gacccgggcc gccgaggccg
     2041 gcgtaaaaga cgtggttttg aagccgcttc tcgacgaaaa cctgctcaag cgtatccgcc
     2101 gcgccatcca ggaccggcct cgggcatgac ctacggggtt ctacgtaagg cacccccctt
     2161 aagatatcgc tcgaaatttt cgaacctccc gataccgcgt accaatgcgt catcacaacg
     2221 gagtctagaa aagaggagaa atactagatg agcacaaaaa agaaaccatt aacacaagag
     2281 cagcttgagg acgcacgtcg ccttaaagca atttatgaaa aaaagaaaaa tgaacttggc
     2341 ttatcccagg aatctgtcgc agacaagatg gggatggggc agtcaggcgt tggtgcttta
     2401 tttaatggca tcaatgcatt aaatgcttat aacgccgcat tgcttgcaaa aattctcaaa
     2461 gttagcgttg aagaatttag cccttcaatc gccagagaaa tctacgagat gtatgaagcg
     2521 gttagtatgc agccgtcact tagaagtgag tatgagtacc ctgttttttc tcatgttcag
     2581 gcagggatgt tctcacctga gcttagaacc tttaccaaag gtgatgcgga gagatgggta
     2641 agcacaacca aaaaagccag tgattctgca ttctggcttg aggttgaagg taattccatg
     2701 accgcaccaa caggctccaa gccgagcttt cctgacggaa tgttaattct cgttgaccct
     2761 gagcaggctg ttgagccagg tgatttctgc atagccagac ttgggggtga tgagtttacc
     2821 ttcaagaaac tgatcaggga tagcggtcag gtgtttttac aaccactaaa cccacagtac
     2881 ccaatgatcc catgcaatga gagttgttcc gttgtgggga aagttatcgc tagtcagtgg
     2941 cctgaagaga cgtttggcgc tgcaaacgac gaaaactacg ctttagtagc ttaataacgc
     3001 tgatagtgct agtgtagatc gctactagag ccaggcatca aataaaacga aaggctcagt
     3061 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     3121 acactggctc accttcgggt gggcctttct gcgtttatat actagagtaa caccgtgcgt
     3181 gttgactatt ttacctctgg cggtgataat ggttgctcta gaaataattt tgtttaactt
     3241 taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc agcggcctgg
     3301 tgccgcgcgg cagccatatg gctagcatga ctggtggaca gcaaatgggt cggatccaaa
     3361 gaggagaaat actagatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg
     3421 gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt
     3481 gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta
     3541 ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg
     3601 gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa
     3661 cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa
     3721 ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat
     3781 atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat
     3841 aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc
     3901 gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg
     3961 ccagacaatc actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc
     4021 gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa
     4081 ctgtacaaat gatgactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     4141 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     4201 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     4261 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4321 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4381 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4441 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4501 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4561 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4621 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4681 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4741 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4801 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     4861 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     4921 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     4981 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     5041 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     5101 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     5161 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     5221 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     5281 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     5341 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     5401 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     5461 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     5521 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     5581 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     5641 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     5701 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5761 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5821 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5881 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     5941 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6001 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6061 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6121 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6181 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6241 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6301 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6361 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6421 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6481 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     6541 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     6601 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     6661 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     6721 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     6781 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     6841 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     6901 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     6961 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     7021 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     7081 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     7141 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     7201 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     7261 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     7321 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     7381 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     7441 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     7501 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     7561 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     7621 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     7681 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     7741 gagctaactt acatcaattg tcaatcgttg agcatgccgg cgcgcatcgc gaggcgaacc
     7801 agctccgaaa ggctgttggc ctgcatcttg gtcatgacgt tggcccgata cacctcgatg
     7861 gtgcgcgggc tgatgtcgta ctcgcgggcg atcagcttgt tggaaaggcc ggc
//