LOCUS       sequence-374346-        6157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Abi1 fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  AUTHORS   Innocenti M, Zucconi A, Disanza A, Frittoli E, Areces LB, Steffen A,
            Stradal TE, Di Fiore PP, Carlier MF, Scita G
  TITLE     Abi1 is essential for the formation and activation of a WAVE2 
            signalling complex.
  JOURNAL   Nat Cell Biol. 2004 Apr;6(4):319-27. Epub 2004 Mar 28.
  PUBMED    15048123
REFERENCE   2  (bases 1 to 6157)
  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: "Nat Cell 
            Biol."; date: "2004-04"; volume: "6(4)"; pages: "319-27. Epub 2004 
            Mar 28"
COMMENT     Sequence Label: pEGFP Abi1
FEATURES             Location/Qualifiers
     source          1..6157
                     /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    2945..3066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2982..3001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3036..3055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3073..3528)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3210..3231)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3422..3441
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3555..3659
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3661..4018
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3661..3852
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3684..3704)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3869..4004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4053..4847
                     /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             4053..4847
                     /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(4107..4126)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4717..4736
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5035..5054)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5079..5126
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5455..6043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5944..5963
                     /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 atctatggca gagctgcaga tgttactaga ggaggagatc
     1381 ccgtctggca agagggcgct gatcgagagt taccagaacc tgactcgggt ggcagactac
     1441 tgtgaaaaca actacataca ggctacagac aagagaaaag ctttagagga gaccaaagcc
     1501 tatacaaccc aatctctagc tagtgttgct tatcaaataa atgcattggc caacaatgta
     1561 ctccagttgc tggatatcca agcctctcag cttcggagaa tggagtcttc catcaatcat
     1621 atctcacaga ctgtggatat tcataaggag aaagtggcac gaagagagat tggtattttg
     1681 acaacaaata agaatacatc aagaactcac aaaataatag cacctgcgaa tatggagcgc
     1741 cctgtaaggt atattcggaa acctatcgat tacacagttc tggatgatgt gggccatggt
     1801 gtcaagtggc taaaagccaa gcatggaaat aaccagcctg caagaactgg cacactgtcg
     1861 agaacaaatc ctcctactca gaaaccgcca agtcctccca tgtcaggccg gggaacactg
     1921 ggacggaata ctccttataa aaccctggaa cctgttaaac ccccaacagt tcctaatgac
     1981 tatatgacca gtcctgctag gcttggaagt cagcatagtc caggcaggac agcatcttta
     2041 aatcagagac caaggacaca cagtggaagt agtggaggaa gtggaagtcg agaaaacagt
     2101 ggtagcagta gtattggcat tcccattgct gtgcctacac cttcgccacc cactattgga
     2161 ccagccccgg gctcagctcc tggttcccag tatggcacaa tgaccaggca gatatctcga
     2221 cacaactcta ctacttcttc gacatcttct ggtggataca gacgaactcc ctctgtgact
     2281 gctcaatttt ctgctcagcc tcatgttaat ggaggtccac tttattctca aaattcaatt
     2341 tctattgctc caccccctcc ccctatgcct cagttgactc cacagatacc tctcacaggc
     2401 ttcgtggcca gggtgcagga aaacattgct gatagtccaa ctccaccgcc accacctcca
     2461 ccagatgaca ttcccatgtt tgatgactct ccacctcccc caccaccacc accagtggat
     2521 tatgaagatg aggaggctgc agtagttcag tataatgatc catatgcaga tggggatcct
     2581 gcttgggccc ccaagaatta tattgagaaa gttgttgcaa tatatgatta tacaaaagac
     2641 aaggatgatg agctgtcatt tatggagggt gcaatcattt atgttataaa gaagaatgat
     2701 gatggctggt atgaaggagt ctgcaatcga gtgactggtc tgttccctgg gaactatgtt
     2761 gaatcaatca tgcactatac tgattaagcg gccgctcgac ggtaccgcgg gcccgggatc
     2821 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2881 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2941 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3001 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3061 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3121 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3181 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3241 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3301 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3361 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3421 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3481 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3541 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3601 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3661 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3721 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3781 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3841 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3901 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3961 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4021 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4081 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4141 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4201 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4261 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4321 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4381 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4441 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4501 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4561 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4621 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4681 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4741 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4801 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4861 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4921 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4981 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5041 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5101 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5161 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5221 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5281 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5341 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5401 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5461 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5521 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5581 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5641 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5701 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5761 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5821 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5881 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5941 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6001 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6061 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6121 ccctgattct gtggataacc gtattaccgc catgcat
//
