LOCUS       sequence-252747-        7836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the N-terminus + central domain of bovine 
            SV1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  AUTHORS   Wulfkuhle JD, Donina IE, Stark NH, Pope RK, Pestonjamasp KN, 
            Niswonger ML, Luna EJ
  TITLE     Domain analysis of supervillin, an F-actin bundling plasma membrane 
            protein with functional nuclear localization signals.
  JOURNAL   J Cell Sci. 1999 Jul:112 (Pt 13):2125-36. doi: 
            10.1242/jcs.112.13.2125
  PUBMED    10362542
REFERENCE   2  (bases 1 to 7836)
  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: "J Cell Sci.
            1999 Jul:112 (Pt 13):2125-36. doi: 10.1242/jcs.112.13.2125"
COMMENT     Sequence Label: EGFP-bSV1_1-1009
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        424..627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     578..598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(672..674,675..677,678..1388)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(672..674,675..677,678..1388)
                     /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(717..738)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(978..997)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1325..1346
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4683..4804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4720..4739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4774..4793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4811..5266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4948..4969)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5160..5179
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5293..5397
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5399..5756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5399..5590
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5422..5442)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5607..5742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5791..6585
                     /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             5791..6585
                     /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(5845..5864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6455..6474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6773..6792)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6817..6864
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7193..7781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7682..7701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
       61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      601 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      661 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      721 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      781 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      841 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      901 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      961 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1021 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1081 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1141 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1201 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1261 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1321 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1381 tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtacct
     1441 tcgatgctca gcttttagct gcccttccta aagttgcaga attaagaaaa atctttgaac
     1501 caaagaggaa agaattttta gaaatgaaaa gaaaagaaag aattgcccgg cgcttagaag
     1561 gaattgaaac cgacacgcag cccatcctct tgcagagctg cacgggcttg gtgactcatc
     1621 gcctgctgga ggaggacaca cccagataca tgcgtgccac agacccggcc agcccgcaca
     1681 ttggtcgatc aaatgaagaa gaagaaactt cagattcttc actagaaaag cagactcgat
     1741 ccaaacagtg cacagaaacc tcaggcatcc atgctgactc gccctacagt tcaggcatca
     1801 tggacaccca gagcctcgag tccaaagccg aaagaatcgc caggtacaaa gcagagagaa
     1861 gacggcagct ggcagaaaag tatgggctca ccctggatcc ggaagcagac tctgaaactc
     1921 catctcgata cagcaggtcc agaaaggacc ccgaggctgc ggagaaaagg ggagtaagaa
     1981 gcgagcgatc ggccgagtcc agcagagacg ccggctcatc ctactccagg actgagctct
     2041 cggggctcag gacctgcgtg gctgagtcca aggactacgg cctgcaccgg agtgacggtg
     2101 tttccgacac agaggtgctg ctcaatgcag aaaaccaaag acgcggtcaa gagcccagtg
     2161 ccaccgggct ggcccgagac ctgccccttg caggggaggt ctcctcttct ttctcattct
     2221 ccgggcgaga ctctgccctc ggtgaagtgc caaggtcccc aaaggcagtg cacagcctgc
     2281 ccagtccatc gcctgggcag ccggcctccc caagccactc caccagtgac ctgccactcc
     2341 ctgcggaggc tcgagccagg tcaacttcaa attcagaaat gccagctgcc gaggatgaag
     2401 aaaaggtcga tgagcgagcg aggctgagcg tcgctgctaa gaggctgctc ttcagggaga
     2461 tggagaagtc gtttgacgag aagagcgtcc caaagcgccg ctcccgcaat gcggccgtgg
     2521 agcagaggct gcggcgcctc caggaccggt cccacacgca gcccgtcacc accgaggagg
     2581 tggtcattgc agccacattg caggcgtctg ctcaccaaaa ggcactagcc agagaccaga
     2641 caaacgagag caaagattct gctgagcagg gagaacccga ctcctccact ctgagcttag
     2701 cggagaagct ggccttgttt aacaaactat ctcagccggt ctcaaaagcc atttcaacac
     2761 ggaacagatt agacatgaga cagaggagga tgaatgctcg ttatcaaacc cagccggtca
     2821 cgcttggaga agtggaacag gtgcagagtg gcaagctcat ggctttctct cccaccatta
     2881 acacttccgt gtccaccgtg gcatctaccg tcccccccat gtatgcagga aatcttcgga
     2941 caaagccact tccagatgac agctttggtg ccactgagca gaagtttgct tcttcgctag
     3001 aaaactccga ctccccagtt agaagcatcc tgaaatccca aggctggcag ccctcggtcg
     3061 agggtgctgg gagcaaagca atgttgagag aatttgaaga gacagaacgc aagggaggct
     3121 tgacaggtgg agatggcggg gttacaaagt atgggtcctt cgaggaagca gagctgtcct
     3181 accccgtcct cagcagagtc cgggagggag acaaccataa agaggccatc tatgcccttc
     3241 cgaggaaagg aagcctggag ctcgcccatc ctcccatcgc ccagcttggt gatgacctga
     3301 aggaattttc caccccaaag agcaccatgc aggcaagccc agactggaag gagaggcagc
     3361 tctttgaaga gaaggtggac ttggagaatg tcacaaagag gaagttttcg ctgaaagccg
     3421 cggagttcgg ggaacccact tctgagcaga ccggcgcggc tgctgggaaa ccggctgctc
     3481 cgactgcaac cccggtgtcc tggaagccgc aggatccctc cgaacagcct caggagaagc
     3541 ggtatcagag cccgtgtgcg atgtttgctg ctggagagat caaagccccg gcggtggagg
     3601 gcagcctcga ctcccccagc aaaaccatgt ccattaaaga aagattagca ctgttgaaga
     3661 agagcggtga ggaagactgg agaaacagac tcaacagaaa gcaggaatac ggcaaagcct
     3721 ccatcaccag cagcctgcac atccaagaga cggagcagtc actcaagaag aagcgggtca
     3781 cagaaagtcg agagagccag atgactattg aagagagaaa gcatctcatc accgtgagag
     3841 aggacgcctg gaagaccagg ggcaaaggag cagccaacga ctctacccag tttactgtgg
     3901 ccggcaggat ggtgaagcga ggtctggcgt cccccactgc catcaccccg gtagcatccc
     3961 ctgtttccag caaagcaagg ggcactacac cagtttccag acccctggaa gatatcgaag
     4021 ccagaccaga catgcagtta gagtcggacc tcaagctgga caggctggaa accttcctaa
     4081 gaaggctgaa taacaaagtt ggtgggatgc aagaaacagt cctcactgtc actgggaaat
     4141 cggtgaaaga ggtgatgaag ccggacgatg atgaaacctt tgccaagttt taccgcagcg
     4201 tggactcctc tctaccgaga agccccgtag agctggacga ggatttcgat gtcatttttg
     4261 atccttatgc ccccaggttg acatcctcgg tggctgagca caagcgtgcc gtgcggccca
     4321 agcgccgggt ccaggcttcc aaaaaccccc tgaagatgct ggccgcgagg gaagatctcc
     4381 tgcaggagta cactgagcag agactgaacg tggccttcgt ggagtcaaag cggatgaaag
     4441 tcgaaaagtt gtccgccaac tccagcttct cagaagtcac cctggcgggg ttagccagca
     4501 aagaaaactt cagcaacgtc agcctgcgga gcgtcaacct gacggaacag aattgatagt
     4561 aagcctctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     4621 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     4681 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4741 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4801 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4861 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4921 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4981 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     5041 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     5101 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     5161 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     5221 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     5281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     5401 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5581 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5641 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5701 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     5761 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5821 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5881 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5941 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     6001 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6061 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6121 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6181 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6241 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6301 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6361 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6421 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6481 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6541 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     6601 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     6661 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     6721 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     6781 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     6841 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6901 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6961 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     7021 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     7081 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7141 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7201 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7261 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7321 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7381 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7441 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7501 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7561 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7621 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7681 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7741 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7801 ttttacggtt cctggccttt tgctggcctt ttgctc
//
