LOCUS       sequence-432429-        6937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mamallian Expression of SV2A GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6937)
  AUTHORS   Dong M, Yeh F, Tepp WH, Dean C, Johnson EA, Janz R, Chapman ER
  TITLE     SV2 is the protein receptor for botulinum neurotoxin A.
  JOURNAL   Science. 2006 Apr 28;312(5773):592-6. doi: 10.1126/science.1123654. 
            Epub 2006 Mar 16.
  PUBMED    16543415
REFERENCE   2  (bases 1 to 6937)
  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"; doi: 
            "10.1126/science.1123654"; journalName: "Science"; date: 
            "2006-04-28- 28"; volume: "312"; issue: "5773"; pages: "592-6"
COMMENT     Sequence Label: pEGFP SV2A GFP
FEATURES             Location/Qualifiers
     source          1..6937
                     /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      2877..2886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2883..2885,2886..2888,2889..3602)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2883..2885,2886..2888,2889..3599)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2928..2949)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3189..3208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3536..3557
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3725..3846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3762..3781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3816..3835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3853..4308)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3990..4011)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4202..4221
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4335..4439
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4441..4798
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4441..4632
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4464..4484)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4649..4784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4833..5627
                     /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             4833..5627
                     /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(4887..4906)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5497..5516
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5815..5834)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5859..5906
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6235..6823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6724..6743
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctatgga agaaggcttt cgagaccgag
      661 cagcgttcat ccgtggggcc aaagacattg ccaaggaagt taagaagcac gcggccaaga
      721 aggtggtgaa gggtctcgac agagtccagg atgaatattt ccgaaggtcc tactcccgct
      781 ttgaggagga ggaggatgat gatgacttcc ctgcccctgc tgacggctat taccgcggag
      841 aaggggccca ggatgaggag gaaggtggcg cttccagtga tgccactgag ggccacgatg
      901 aggatgatga gatctacgag ggagaatatc agggcatccc ccgggcagag tctgggggca
      961 aaggcgaacg gatggcagat ggggcacccc tggctggagt gagagggggc ttaagtgatg
     1021 gggagggtcc ccctgggggt cgcggggagg cgcagcggcg taaagatcgg gaagaattgg
     1081 ctcagcagta tgagaccatc ctccgggagt gcggccatgg tcgcttccag tggacactct
     1141 acttcgtgct gggtctggcg ctgatggccg atggtgtaga ggtctttgtg gtgggctttg
     1201 tgctgcccag tgctgagaaa gatatgtgcc tgtcggactc caacaaaggc atgctaggcc
     1261 tcattgtgta cctgggcatg atggtggggg ccttcctctg gggaggcctg gctgatcggc
     1321 tgggtcggag acagtgtctg ctcatctcgc tctcagtcaa cagcgtcttc gccttcttct
     1381 catccttcgt ccagggttat ggcaccttcc ttttctgccg cctcctttct ggggttggga
     1441 ttggtggttc catccccatt gtcttctcct atttttcgga gtttctggcc caggagaaac
     1501 gtggggagca tttgagctgg ctctgtatgt tctggatgat tggtggcgtg tatgcagctg
     1561 caatggcctg ggccatcatc ccccactatg ggtggagttt ccagatgggc tctgcttacc
     1621 agttccacag ctggagggtc tttgtcctcg tctgtgcctt tccctctgtg tttgccatcg
     1681 gggctctgac tacgcagccg gagagtcccc gcttcttctt agagaatggg aagcacgatg
     1741 aggcctggat ggtgctgaag caggttcatg acaccaacat gcgagccaag ggccatcctg
     1801 agcgagtctt ctcagtaacc cacattaaaa cgattcatca ggaggatgaa ttgattgaga
     1861 tccagtcaga cacaggaacc tggtaccagc gctggggagt gcgggctttg agcctggggg
     1921 gtcaggtttg ggggaacttc ctctcctgct tcagtccaga gtaccggcgc atcactctga
     1981 tgatgatggg ggtatggttc accatgtcct tcagctacta cggtttgact gtctggtttc
     2041 ccgacatgat ccgccatctc caggctgtgg actatgcagc ccgaaccaaa gtgttcccag
     2101 gggagcgcgt ggagcacgtg acatttaact tcacactgga gaatcagatc caccgagggg
     2161 gacagtactt caatgacaag ttcatcgggc tgcgtctgaa gtcagtgtcc tttgaggatt
     2221 ccctgtttga ggaatgttac tttgaagatg tcacatccag caacacattc ttccgcaact
     2281 gcacattcat caacaccgtg ttctacaaca cggacctgtt tgagtacaag ttcgtgaaca
     2341 gccgcctggt gaacagcaca ttcctgcaca ataaggaagg ttgcccacta gatgtgacag
     2401 ggacgggcga aggtgcctac atggtgtact ttgtcagctt cttggggaca ctggctgtgc
     2461 tccctggaaa tattgtgtct gctctgctca tggacaagat tggcaggctc agaatgcttg
     2521 ctggttccag tgtgttgtcc tgtgtttcct gcttcttcct gtcttttggg aacagtgagt
     2581 cagccatgat cgctctgctc tgcctttttg ggggagtcag tattgcatcc tggaacgcgc
     2641 tggacgtgct gactgttgaa ctctaccctt ccgacaagag gacgacggcc ttcggcttcc
     2701 tgaatgccct gtgtaagctg gcagctgtac tgggcatcag catcttcacg tcctttgtgg
     2761 gaatcaccaa ggccgctccc atcctcttcg cctcagctgc gcttgccctt ggtagctctc
     2821 tggctctgaa gctgcctgag acccggggac aggtgctgca gcgggatcca ccggtcgcca
     2881 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2941 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3001 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3061 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3121 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3181 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3241 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3301 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3361 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3421 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3481 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3541 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3601 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3661 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3721 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3781 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3841 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3901 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3961 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4021 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4081 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4141 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4201 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4261 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4321 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4381 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4441 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4501 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4621 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4681 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4741 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4801 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4861 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4921 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4981 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5041 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5101 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5161 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5221 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5281 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5341 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5401 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5461 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5521 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5581 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5641 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5701 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5761 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5821 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5881 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5941 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6001 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6061 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6121 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6181 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6241 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6301 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6361 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6421 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6481 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6541 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6601 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6661 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6721 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6781 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6841 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6901 ccctgattct gtggataacc gtattaccgc catgcat
//
