LOCUS Lact-C2-GFP 5194 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Lact-C2-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5194)
AUTHORS Yeung T, Gilbert GE, Shi J, Silvius J, Kapus A, Grinstein S
TITLE Membrane phosphatidylserine regulates surface charge and protein
localization.
JOURNAL Science. 2008 Jan 11. 319(5860):210-3.
PUBMED 18187657
REFERENCE 2 (bases 1 to 5194)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..5194
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 115..418
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 419..622
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 573..593
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 661..670
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 667..1383
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 667..1383
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(712..733)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(973..992)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1320..1341
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2036..2157
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2073..2092)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2127..2146
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2164..2619)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2301..2322)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2513..2532
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2646..2750
/gene="bla"
/label=AmpR promoter
promoter 2752..3109
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2775..2795)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2960..3095
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3022..3041
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3144..3938
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3198..3217)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3808..3827
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4126..4145)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4170..4217
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4546..5134
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5035..5054
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
661 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
721 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
781 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
841 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
901 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
961 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
1021 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
1081 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
1141 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
1201 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
1261 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
1321 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
1381 aagtccggac tcagatcttg cactgaaccc ctaggcctga aggataatac catccccaac
1441 aagcagatca cagcctccag ctactacaaa acctggggcc tgagtgcctt tagctggttt
1501 ccctactacg cacgactgga taatcagggc aagttcaacg cctggaccgc ccagaccaac
1561 agtgcctctg agtggctgca gattgacctg ggctcccaga agcgagtcac gggcatcatc
1621 acccagggtg cccgagactt tggccacatt caatatgtgg ctgcctacag ggtggcctat
1681 ggtgatgatg gtgtgacctg gactgagtac aaggacccgg gggcctcaga aagcaagatt
1741 ttccctggta acatggacaa taattcccac aagaagaaca tatttgagac gccgttccag
1801 gctcgcttcg tgcggatcca gcccgtggcc tggcacaacc gtatcaccct gcgagtggag
1861 ctgctgggct gttaggaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
1921 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
1981 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
2041 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
2101 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
2161 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
2221 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
2281 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
2341 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
2401 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
2461 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
2521 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
2581 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
2641 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
2701 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
2761 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
2821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
2941 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
3001 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
3061 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
3121 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
3181 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
3241 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
3301 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
3361 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
3421 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3481 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3541 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
3601 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
3661 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
3721 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
3781 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3841 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3901 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
3961 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
4021 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
4081 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
4141 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
4201 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
4261 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
4321 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
4381 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
4441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
4501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
4561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
4621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
4681 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
4741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
4801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
4861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
4921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
4981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
5041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
5101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
5161 gttcctggcc ttttgctggc cttttgctca catg
//
