LOCUS CMV-SEAP 6034 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS CMV-SEAP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6034)
TITLE pLAI-Delta-mls
REFERENCE 2 (bases 1 to 6034)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..6034
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(35..1552)
/codon_start=1
/product="secreted alkaline phosphatase from human
placenta"
/label=SEAP
/note="The natural enzyme has a C-terminal transmembrane
sequence that is replaced by a GPI anchor."
/translation="MLGPCMLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAA
KKLQPAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKT
YNVDKHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGK
SVGVVTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVIL
GGGRKYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLD
PSVTHLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHG
HHESRAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGL
APGKARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGED
VAVFARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
promoter complement(1717..1920)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(1746..1766)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 1921..2300
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 2402..2421
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(2533..2552)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2534..2552)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2559..2576)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2559..2575)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2568..2590)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2717..3169
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2801..2820)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3011..3032
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3195..3299
/gene="bla"
/label=AmpR promoter
CDS 3300..4160
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3518..3537)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4331..4919
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4820..4839
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5073..5090
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5207..5228
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5243..5273
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5281..5297
/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 5286..5308
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5305..5321
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5305..5321
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 5342..5360
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 5406..5540
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5430..5449)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5484..5503
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
ORIGIN
1 acgcggggac cacgggttaa cccgggtgcg cggcgtcggt ggtgccggcg gggggcgcca
61 ggtcgcaggc ggtgtagggc tccaggcagg cggcgaaggc catgacgtgc gctatgaagg
121 tctgctcctg cacgccgtga accaggtgcg cctgcgggcc gcgcgcgaac accgccacgt
181 cctcgcctgc gtgggtctct tcgtccaggg gcactgctga ctgctgccga tactcggggc
241 tcccgctctc gctctcggta acatccggcc gggcgccgtc cttgagcaca tagcctggac
301 cgtttccgta taggaggacc gtgtaggcct tcctgtcccg ggccttgcca ggggccagcc
361 cgaagatgga gctccctcgc agggggtagc ctccgaagga gaagacgtgg gagtggtcgg
421 cagtgacgag gctcagcgtg tcctcctcgc tggtgagctg gcccgccctc tcaatggcgt
481 cgtcgaacat gatcgtctca gtcagtgccc ggtaagccct gctttcatga tgaccatggt
541 cgatgcgacc accctccacg aagaggaaga agccgcgggg gttcctgctc agcaggcgca
601 gggcagcctc tgtcatctcc atcagggagg ggtccagtgt ggagtctcgg tggatctcgt
661 atttcatgtc tccaggctca aagagaccca tgagatgggt cacagacggg tccagggaag
721 cctgcatgag ctcagtgcgg ttccacacat accgggcacc ctggcgcttc gccagccatt
781 cctgcaccag attcttcccg tccagcctgg tcccaccttg gctgtagtca tctgggtact
841 cagggtctgg ggttcccatg cgaaacatgt actttcggcc tccacctagg atcacgtcaa
901 tgtccatgtt ggagatgagc tgcgtagcga tgtcctggca cccctcctgg cgggccgagg
961 caggcacgtc ggcgtccgag taccagttgc ggttcaccgt gtgggcgtag gtgccggctg
1021 gcgaggcgtg ctgcactcgt gtggtggtta ccactcccac tgacttccct gctttcttgg
1081 cccgattcat cacggagatg acctcgttgc cgcgtgtcgt gttgcactgg ttaaagcggg
1141 cggctgcact caagccaatg gtctggaagt tgcccttgac cccgcacagg taggccgtgg
1201 ctgtggctcc actgtctggc acatgtttgt ctacattgta tgtcttggac agagccacat
1261 atgggaagcg gtccatggcc aggggtatct caggccccag tttgtccttc ttctgccctt
1321 ttaggatcct ggcagctgtc accgtagaca cccccatccc atcgcccagg aagatgatga
1381 ggttcttggc ggctgtctgt gcaggctgca gcttcttggc ggcacccagg gcctcggctg
1441 cctcgcggtt ccagaagtcc gggttctcct cctcaactgg gatgatgccc agggagagct
1501 gtagcctcag gcccagcagc agcagcagca gcagcatgca gggccccagc atgtctggag
1561 ggcagcagga cagtggcgag gcaggaatta gcttgaggtg cacaccaatg tggtgaatgg
1621 tcaaatggcg tttattgtat cgagctaggc acttaaatac aatatctctg caatgcggaa
1681 ttcgatatca agcttatcga tacggttcac taaacgagct ctgcttatat agacctccca
1741 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
1801 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
1861 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
1921 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
1981 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
2041 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
2101 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
2161 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
2221 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
2281 actagtcaat aatcaatgtc aacatggcgg taatgttgga catgagccaa tataaatgta
2341 catattatga tatggataca acgtatgcaa tgggccaagc tcatggctga ctaatttttt
2401 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
2461 gcttttttgg aggcctaggc ttttgcaaaa agctcgacgc gaggctcgag ggggggcccg
2521 gtacccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
2581 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
2641 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
2701 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggttacg
2761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
2821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
2881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
2941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
3001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
3061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
3121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
3181 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
3241 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
3301 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
3361 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
3421 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
3481 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
3541 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
3601 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
3661 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
3721 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
3781 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
3841 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
3901 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
3961 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
4021 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
4081 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
4141 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
4201 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
4261 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
4321 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
4381 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
4441 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
4501 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
4561 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
4621 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
4681 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
4741 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
4801 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
4861 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
4921 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
4981 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
5041 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
5101 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
5161 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
5221 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
5281 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg
5341 aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctcta
5401 ggatcgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
5461 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
5521 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
5581 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt
5641 atgatcaatg aattgctcta gaggatcccc gggtacccac tgggctgtag gtggtggctg
5701 gatggcctga gccccctcct ttcctggggt gcaggcagcc acagggcaca acctgcagtc
5761 cctggggttg gggtagggag tccgagagca gccaaggggc aggtttggtg tcatgggctc
5821 agtccccaga gcgcacgggg aggggaaggt tccatggcag gacctctgta tagggactcc
5881 ggggatgacg actcgggctg gaggccaggc aggacgacag gaggtgggga gcaggagaac
5941 tccgggatgg ggaagcagga gccccaggga cgggacactc agggagcagt ggccgtctcc
6001 agcagcagca gggtcccggc cagcagagga agca
//
