LOCUS       sequence-51352-e        6094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6094)
  AUTHORS   An SJ, Almers W
  TITLE     Tracking SNARE complex formation in live endocrine cells.
  JOURNAL   Science. 2004 Nov 5;306(5698):1042-6.
  PUBMED    15528447
REFERENCE   2  (bases 1 to 6094)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2004-11-5"; volume: "306(5698)"; pages: "1042-6"
COMMENT     Sequence Label: SCORE
FEATURES             Location/Qualifiers
     source          1..6094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(612..614,615..617,618..1328)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1836..1845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1842..1844,1845..1847,1848..2555)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     polyA_signal    2882..3003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2919..2938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2973..2992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3010..3465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3147..3168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3359..3378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3492..3596
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3598..3955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3598..3789
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3621..3641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3806..3941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3990..4784
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3990..4784
                     /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(4044..4063)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4654..4673
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4972..4991)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5016..5063
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5392..5980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5881..5900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgctc
      601 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      661 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      721 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      781 cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc cgctaccccg
      841 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      901 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      961 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1021 tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc accgccgaca
     1081 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg
     1141 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1201 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1261 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1321 tgtacaagtc cggaggttct ggtggttctg gtggttctgg tggttctggt atggccgaag
     1381 acgcagacat gcgcaatgag ctggaggaga tgcagaggag ggctgaccag ctggctgatg
     1441 agtccctgga aagcacccgt cgcatgctgc agctggttga agagagtaaa gatgctggca
     1501 tcaggacttt ggttatgttg gatgagcaag gcgaacaact ggaacgcatt gaggaaggga
     1561 tggaccaaat caataaggac atgaaagaag cagaaaagaa tttgacggac ctaggaaaat
     1621 tctgcgggct ttgtgtgtgt ccctgtaaca agcttaaatc cagtgatgct tacaaaaaag
     1681 cctggggcaa taatcaggat ggagtagtgg ccagccagcc tgcccgtgtg gtggatgaac
     1741 gggagcagat ggccatcagt ggtggcttca tccgcagggt aacaaatgat gccggtggtt
     1801 ctggtggttc tggtggttct ggtggttcac cggtcgccac catggtgagc aagggcgagg
     1861 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1921 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     1981 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct
     2041 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2101 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2161 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2221 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2281 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     2341 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     2401 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     2461 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2521 ccgccgggat cactctcggc atggacgagc tgtacagtgg tggttctggt ggttctcgag
     2581 aaaatgaaat ggatgaaaac ctagagcagg tgagcggcat catcggaaac ctccgccata
     2641 tggctctaga catgggcaat gagattgaca cccagaatcg ccagatcgac aggatcatgg
     2701 agaaggctga ttccaacaaa accagaattg atgaagccaa ccaacgtgaa acaaagatgc
     2761 tgggaagtgg ttaatgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2821 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2881 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2941 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3001 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3061 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3121 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3181 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3241 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3301 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3361 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3421 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3601 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3961 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4021 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4081 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4141 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4201 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4261 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4321 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4381 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4441 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4501 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4561 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4621 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4681 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4741 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4801 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4861 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4921 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4981 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5041 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5101 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5161 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5221 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5281 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5341 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5521 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6061 tgattctgtg gataaccgta ttaccgccat gcat
//
