LOCUS       sequence-381067-        6236 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 6236)
  AUTHORS   Hussain NK, Jenna S, Glogauer M, Quinn CC, Wasiak S, Guipponi M, 
            Antonarakis SE, Kay BK, Stossel TP, Lamarche-Vane N, McPherson PS
  TITLE     Endocytic protein intersectin-l regulates actin assembly via Cdc42 
            and N-WASP.
  JOURNAL   Nat Cell Biol. 2001 Oct;3(10):927-32.
  PUBMED    11584276
REFERENCE   2  (bases 1 to 6236)
  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: "2001-10"; volume: "3(10)"; pages: "927-32"
COMMENT     Sequence Label: GFP-N-WASP
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     155..174
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     414..435
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      477..486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(525..728)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(554..574)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        729..1032
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1207..1795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1287..1306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2124..2171
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2196..2215
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2403..3197)
                     /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             complement(2403..3197)
                     /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(2514..2533)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3124..3143
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3232..3589)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3246..3381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3398..3589
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3546..3566
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3591..3695)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3722..4177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3809..3828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4019..4040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4184..4305
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4195..4214)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4249..4268
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6000..6236,1..474,475..477,478..480))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(6000..6236,1..474,475..477,478..480))
                     /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(6042..6063)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
       61 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
      121 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
      181 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
      241 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
      301 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
      361 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
      421 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
      481 ggtggcgacc ggtagcgcta gcggatctga cggttcacta aaccagctct gcttatatag
      541 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
      601 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
      661 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
      721 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
      781 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
      841 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
      901 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
      961 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     1021 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     1081 tattaataac taatgcatgg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1921 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc
     1981 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga
     2041 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc
     2101 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct
     2161 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     2221 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     2281 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     2341 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     2401 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     2461 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     2521 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     2581 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     2641 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc
     2701 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     2761 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     2821 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     2881 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     2941 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     3001 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     3061 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     3121 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     3181 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     3241 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     3301 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     3361 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     3421 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     3481 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     3541 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     3601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3721 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3841 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4081 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4141 aacgcgaatt ttaacaaaat attaacgctt acaatttacg cgttaagata cattgatgag
     4201 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4261 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4321 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     4381 ctctacaaat gtggtatggc tgattatgat cagttatcta gatccggtgg atcccgggcc
     4441 cgcggtaccg tcgactgcag aattcgaagc tttcagtctt cccactcatc atcatcctca
     4501 aaatcttctt catcatcatc atcttcatct tcatctgagg aatgaatggc tttgctcctt
     4561 ttctgcatca cttccatcag cgcacccaca attcctgaag tgggcgcggg ggttggtggt
     4621 gtggactctt ggccatcaga cacggatttc aactgaatgc cctgtcgtat ctggtctaga
     4681 agtgcatccc ttcctgagca ggacacgggc cgactattct gctccacttt ttttagctga
     4741 gcaccctctc taatttgatc caaaagagct gctttgtttc ctgaagaagc tggaacttga
     4801 tggtcaccat cagaaggcag gccaggggga ggtggtggcc ctggtggagg gggaggtggt
     4861 ggaggaggtg gtggtgctgt ggaccctgcc atagacagag gcggaggtgg tggtgggcct
     4921 gaaggtgctg aggaaggcag ggctggtggt ggaggagggt acatcctgtt tggaggaggt
     4981 ggaggaacga caacaccagg cctagaagga ggtggaggtg gaggtgcagc agtaggagct
     5041 cttgatggtg gcgggggagg agcccccctt ccacgggcag gagggggagg agggcctgag
     5101 ctgtgaggag ggggaggagg agggggaggt cctcctcttg agggtggagg aggtggtggt
     5161 gcttgccttc ggagttcatt tttaacagct tctacacctc ctgttttttc aataaagtca
     5221 taaataactt ttgatgtttc tctgtcttta agctgggcct cagagatccc acacatatca
     5281 aaaagattct tcaattctgg atccaaatta tttagatcaa aacctgtatt tggatcccaa
     5341 ccaacatgtc caatgtgctg gaaattactt ggtgttccaa tatctgcctt ggttaatctc
     5401 ttctttttag cttttccttt cttcttttct ttggtgtggg agatgttgtt gacttgtgaa
     5461 ctataaaacc tgtttgttgt gatttctgga ttttttatgt caactgtagc catgggtaga
     5521 ttgggaccat ttggagcatc tcgtcttttt tcagatttcc tttgccgtcg acccaacagg
     5581 tctgtaactg cttttcggaa cttttttgct tcttcttcat tggcaaaatt aagagctact
     5641 tgacaagtat ctccagcaaa ggtatgaaaa tatcctctag gactattata tacaaagtta
     5701 ttgtatagct cttgttccca cagtaatttc ccatccttaa tgtcaaatat tcttaaaaaa
     5761 taagatctct gaggattgtc cttaacaaga caagcaacac cactgcactt ctttgaccac
     5821 atacagttcc gatcagctgc atataactgc accactgctg aagacatagt cacacatttc
     5881 ttgccgagga aggagaaaag agactcgttt tcttgcgggg tgagcagcag ggagcccacg
     5941 ttggtgaccc tccgcggggg ctgctggccc gagctcctcg agatctgagt ccggccggac
     6001 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     6061 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     6121 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     6181 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgcc
//
