LOCUS       sequence-456474-        6009 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 6009)
  AUTHORS   Cortesio CL, Perrin BJ, Bennin DA, Huttenlocher A
  TITLE     Actin-binding protein-1 interacts with WASp-interacting protein to 
            regulate growth factor-induced dorsal ruffle formation.
  JOURNAL   Mol Biol Cell. 2010 Jan 1. 21(1):186-97.
  PUBMED    19910490
REFERENCE   2  (bases 1 to 6009)
  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: "Mol Biol 
            Cell. 2010 Jan 1. 21(1):186-97."
COMMENT     Sequence Label: pEGFPc2 mAbp1
FEATURES             Location/Qualifiers
     source          1..6009
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2797..2918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2834..2853)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2888..2907
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2925..3380)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3062..3083)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3274..3293
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3407..3511
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3513..3870
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3513..3704
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3536..3556)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3721..3856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3905..4699
                     /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             3905..4699
                     /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(3959..3978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4569..4588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4887..4906)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4931..4978
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5307..5895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5796..5815
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagctagct tcgaattcat ggcggtgaac
     1381 ctgagccgga acgggccggc gctgcaggag gcctacgtgc gcgtagtcac cgagaaatcc
     1441 ccgaccgact gggctctttt tacctatgaa ggcaacagca atgacatccg tgtggctggc
     1501 acaggagagg gaggcctgga ggagctggtg gaagagctca acagcgggaa ggtgatgtac
     1561 gccttctgca gggtgaagga ccccaactcc ggcctgccca agtttgtcct catcaactgg
     1621 acaggagagg gtgtgaatga tgtgcggaaa ggagcatgtg ccaaccacgt cagcaccatg
     1681 gccaacttcc tgaagggtgc ccacgtgacc atcaatgccc gggccgagga ggatgtggag
     1741 cctgagtgca tcatggagaa ggttgccaag gcctctgggg ccaactacag cttccataag
     1801 gaaagcacct ccttccagga tgtagggccg caggccccag tgggctctgt gtaccagaag
     1861 accaatgcca tatctgagat caagagagtc ggcaaggata acttctgggc caaagctgag
     1921 aaggaagaag agaaccgccg cctggaggag aagcggcgtg ccgaagagga gcggcagcgg
     1981 ttggaggagg agcgacgaga gcgggagctg caggaggctg cccgacgtga gcagcgctac
     2041 caggaacagc acagatcagc tggagccccg aggacaggtg agccagagca ggaagccgtt
     2101 tcaaggacca gacaggagtg ggagtctgct gggcagcagg ccccacaccc acgagagatt
     2161 ttcaagcaga aggaaagggc aatgtccacc acctctgtca ccagctcgca gccgggcaag
     2221 ctgaggagcc ccttcctgca gaagcaactc actcaaccag aaacctccta cggccgagag
     2281 cccacagctc ctgtctcccg gcctgcagca ggtgtctgtg aggagccagc gcctagcact
     2341 ctgtcttctg cccagacaga agaagaacct acatatgaag tacccccaga gcaggacacc
     2401 ctctatgagg aaccaccact ggtacagcag caaggggctg gctccgaaca cattgacaac
     2461 tacatgcaga gccagggctt cagtggacaa gggctgtgcg cccgggcctt gtatgactac
     2521 caggcagctg atgacaccga gatctccttt gaccctgaga acctaatcac aggcatcgag
     2581 gtgattgacg aaggctggtg gcgaggctat gggcctgacg gccactttgg catgtttcct
     2641 gccaactacg tggagctcat agagtgagga tccaccggat ctagataact gatcataatc
     2701 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2761 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2821 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2881 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2941 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3001 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3061 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3121 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3181 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3241 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3301 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3361 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3421 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3481 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3541 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3601 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3721 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3781 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3841 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3901 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3961 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4021 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4081 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4141 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4201 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4261 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4321 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4381 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4441 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4501 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4561 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4621 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4681 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4741 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4801 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4861 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4921 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4981 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5041 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5101 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5161 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5221 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5941 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6001 gccatgcat
//
