LOCUS       sequence-129625-        7095 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 7095)
  AUTHORS   Manabe R, Kovalenko M, Webb DJ, Horwitz AR
  TITLE     GIT1 functions in a motile, multi-molecular signaling complex that 
            regulates protrusive activity and cell migration.
  JOURNAL   J Cell Sci. 2002 Apr 1. 115(Pt 7):1497-510.
  PUBMED    11896197
REFERENCE   2  (bases 1 to 7095)
  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.
            2002 Apr 1. 115(Pt 7):1497-510."
COMMENT     Sequence Label: pEGFP-GIT1
FEATURES             Location/Qualifiers
     source          1..7095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(620..622,623..625,626..1336)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(620..622,623..625,626..1336)
                     /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(665..686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(926..945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1273..1294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3890..4011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3927..3946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3981..4000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4018..4473)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4155..4176)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4367..4386
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4500..4604
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4606..4963
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4606..4797
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4629..4649)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4814..4949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4998..5792
                     /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             4998..5792
                     /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(5052..5071)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5662..5681
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5980..5999)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6024..6071
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6400..6988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6889..6908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga
     1381 catgtcccga aaggggccgc gagcggaggt gtgtgcggac tgcagcgccc cggaccctgg
     1441 ctgggcatcc atcagcaggg gtgtgctggt gtgtgacgag tgctgcagcg tgcaccggag
     1501 cctgggacgc cacatctcca ttgtcaagca ccttcgccac agcgcctggc ctcccacgct
     1561 gctgcagatg gtgcacacgc ttgccagcaa cggggccaac tccatctggg agcactccct
     1621 gctggacccc gcacaagtgc agagcggccg gcgtaaagcc aacccccaag acaaagtcca
     1681 ccccatcaag tcagagttca tcagggccaa gtaccagatg ctggcatttg tgcacaagct
     1741 tccctgccgg gacgatgatg gagtcaccgc caaagacctc agcaagcaac tacactcgag
     1801 cgtgcggaca ggcaacctgg agacatgtct gcgcctgctc tccctgggtg cccaggccaa
     1861 cttcttccac ccagagaagg gcaccacacc tctgcacgtg gctgccaagg caggacagac
     1921 actgcaggcc gagctgcttg tagtgtatgg ggctgaccct ggctcccctg atgttaatgg
     1981 ccgcacaccc attgactatg ccaggcaggc ggggcaccat gagctggcgg aaaggctggt
     2041 tgagtgccaa tatgagctca ctgaccggct ggccttctac ctctgtggac gcaagccgga
     2101 tcacaagaat gggcattaca tcatcccaca gatggctgac agatctcggc aaaagtgcat
     2161 gtctcagagc cttgacttat ccgaattggc caaagctgct aagaagaagc tgcaggcgct
     2221 cagcaaccgg ctttttgagg aactcgccat ggacgtgtat gacgaggtgg atcgaagaga
     2281 aaatgatgca gtgtggctgg ctacccaaaa ccacagcact ctggtgacag agcgcagtgc
     2341 cgtgcccttc ctgcctgtta acccggaata ctcagccacg cggaatcagg ggcgacaaaa
     2401 gctggcccgc tttaatgccc gagagtttgc caccttgatc atcgacattc tcagtgaggc
     2461 caagcggaga cagcagggca agagcctgag cagccccaca gacaacctcg agctgtctct
     2521 gcggagccag agtgacctcg acgaccaaca cgactacgac agcgtggcct ctgacgagga
     2581 cacagaccag gagcccctgc gcagcaccgg cgccactcgg agcaaccggg cccggagcat
     2641 ggactcctcg gacgtgtctg acggggctgt gacgctgcag gagtacctgg agctgaagaa
     2701 ggccttggct acatcggagg caaaggtgca gcagctcatg aaggtcaaca gtagcctgag
     2761 cgacgagctc cggaggctgc agcgagagat ccacaagctg caggcgaaga acctgcagct
     2821 ccggcagcct ccagggccgg tgcccacacc tccactcccc agtgaacggg cggaacacac
     2881 acccatggcg ccaggcggga gcacacaccg cagggatcgc caggcctttt ccatgtatga
     2941 acctggctct gccctgaagc cctttggggg cccccctggg gacgagctca ctacgcggct
     3001 gcagcctttc cacagcactg agctagagga cgacgccatc tattcagtgc acgtccctgc
     3061 tggcctttac cggatccgga aaggggtgtc tgcctcagct gtgcccttca ctccctcctc
     3121 cccgctgccg tcctgctccc aggagggaag ccgccacacg agcaagcttt cccgccacgg
     3181 cagtggagcc gacagtgact atgagaacac gcaaagtggg gacccactgc tggggctgga
     3241 agggaagagg tttctagagc tgggcaaaga ggaagacttc cacccagagc tggaaagcct
     3301 ggatggagac ctagatcctg ggcttcccag cacagaggat gtcatcttga agacagagca
     3361 ggtcaccaag aacattcagg aactgttgcg ggcagcccag gagttcaagc atgacagctt
     3421 cgtgccctgc tcagagaaga tccatttggc tgtgaccgag atggcctccc tcttcccaaa
     3481 gaggccagcc ctggagccag tgcggagctc actgcggctg ctcaacgcca gcgcctaccg
     3541 gctgcagagt gagtgccgga agacagtgcc cccagagccc ggcgccccag tggacttcca
     3601 gctgctgact cagcaggtga tccagtgcgc ctatgacatc gccaaggctg ccaagcagct
     3661 ggtcaccatc accacccgag agaagaagca gtgacctctc tccccacacc ctcacctgca
     3721 ccctagatcc atcacactgg cggccgccac cgcgggcccg ggatccaccg gatctagata
     3781 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3841 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     4021 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4081 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4141 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4201 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4261 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4321 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4381 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4441 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4501 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4561 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4621 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4681 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4741 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4801 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4861 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4921 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4981 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5041 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5101 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5161 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5221 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5281 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5341 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5401 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5461 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5521 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5581 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5641 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5701 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5761 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5821 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5881 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5941 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     6001 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     6061 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6121 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6181 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6241 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6301 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6721 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6781 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6841 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6901 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6961 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7021 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7081 taaccgtatt accgc
//
