LOCUS       sequence-371463-        6294 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 6294)
  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 6294)
  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: GIT1A-FLAG
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2571..2594
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2718..2839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2755..2774)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2809..2828
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2846..3301)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2983..3004)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3195..3214
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3328..3432
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3434..3791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3434..3625
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3457..3477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3642..3777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3826..4620
                     /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             3826..4620
                     /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(3880..3899)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4490..4509
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4808..4827)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4852..4899
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5228..5816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5717..5736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5991..6294
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgacatgtcc cgaaaggggc cgcgagcgga ggtgtgtgcg gactgcagcg
      301 ccccggaccc tggctgggca tccatcagca ggggtgtgct ggtgtgtgac gagtgctgca
      361 gcgtgcaccg gagcctggga cgccacatct ccattgtcaa gcaccttcgc cacagcgcct
      421 ggcctcccac gctgctgcag atggtgcaca cgcttgccag caacggggcc aactccatct
      481 gggagcactc cctgctggac cccgcacaag tgcagagcgg ccggcgtaaa gccaaccccc
      541 aagacaaagt ccaccccatc aagtcagagt tcatcagggc caagtaccag atgctggcat
      601 ttgtgcacaa gcttccctgc cgggacgatg atggagtcac cgccaaagac ctcagcaagc
      661 aactacactc gagcgtgcgg acaggcaacc tggagacatg tctgcgcctg ctctccctgg
      721 gtgcccaggc caacttcttc cacccagaga agggcaccac acctctgcac gtggctgcca
      781 aggcaggaca gacactgcag gccgagctgc ttgtagtgta tggggctgac cctggctccc
      841 ctgatgttaa tggccgcaca cccattgact atgccaggca ggcggggcac catgagctgg
      901 cggaaaggct ggttgagtgc caatatgagc tcactgaccg gctggccttc tacctctgtg
      961 gacgcaagcc ggatcacaag aatgggcatt acatcatccc acagatggct gacagatctc
     1021 ggcaaaagtg catgtctcag agccttgact tatccgaatt ggccaaagct gctaagaaga
     1081 agctgcaggc gctcagcaac cggctttttg aggaactcgc catggacgtg tatgacgagg
     1141 tggatcgaag agaaaatgat gcagtgtggc tggctaccca aaaccacagc actctggtga
     1201 cagagcgcag tgccgtgccc ttcctgcctg ttaacccgga atactcagcc acgcggaatc
     1261 aggggcgaca aaagctggcc cgctttaatg cccgagagtt tgccaccttg atcatcgaca
     1321 ttctcagtga ggccaagcgg agacagcagg gcaagagcct gagcagcccc acagacaacc
     1381 tcgagctgtc tctgcggagc cagagtgacc tcgacgacca acacgactac gacagcgtgg
     1441 cctctgacga ggacacagac caggagcccc tgcgcagcac cggcgccact cggagcaacc
     1501 gggcccggag catggactcc tcggacgtgt ctgacggggc tgtgacgctg caggagtacc
     1561 tggagctgaa gaaggccttg gctacatcgg aggcaaaggt gcagcagctc atgaaggtca
     1621 acagtagcct gagcgacgag ctccggaggc tgcagcgaga gatccacaag ctgcaggcga
     1681 agaacctgca gctccggcag cctccagggc cggtgcccac acctccactc cccagtgaac
     1741 gggcggaaca cacacccatg gcgccaggcg ggagcacaca ccgcagggat cgccaggcct
     1801 tttccatgta tgaacctggc tctgccctga agccctttgg gggcccccct ggggacgagc
     1861 tcactacgcg gctgcagcct ttccacagca ctgagctaga ggacgacgcc atctattcag
     1921 tgcacgtccc tgctggcctt taccggatcc ggaaaggggt gtctgcctca gctgtgccct
     1981 tcactccctc ctccccgctg ccgtcctgct cccaggaggg aagccgccac acgagcaagc
     2041 tttcccgcca cggcagtgga gccgacagtg actatgagaa cacgcaaagt ggggacccac
     2101 tgctggggct ggaagggaag aggtttctag agctgggcaa agaggaagac ttccacccag
     2161 agctggaaag cctggatgga gacctagatc ctgggcttcc cagcacagag gatgtcatct
     2221 tgaagacaga gcaggtcacc aagaacattc aggaactgtt gcgggcagcc caggagttca
     2281 agcatgacag cttcgtgccc tgctcagaga agatccattt ggctgtgacc gagatggcct
     2341 ccctcttccc aaagaggcca gccctggagc cagtgcggag ctcactgcgg ctgctcaacg
     2401 ccagcgccta ccggctgcag agtgagtgcc ggaagacagt gcccccagag cccggcgccc
     2461 cagtggactt ccagctgctg actcagcagg tgatccagtg cgcctatgac atcgccaagg
     2521 ctgccaagca gctggtcacc atcaccaccc gagagaagaa gcagtcgaca gattacaagg
     2581 atgacgacga taagtagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     2641 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2701 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2761 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2821 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2881 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2941 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3001 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3061 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3121 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3181 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3241 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3301 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3361 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3421 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3661 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3721 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3781 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4321 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4381 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4441 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4561 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4621 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4681 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4741 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4801 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4861 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4921 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4981 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5041 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5101 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5161 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5221 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5281 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5341 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5401 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5461 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5521 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5581 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5641 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5701 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5761 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5821 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5881 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     5941 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     6001 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     6061 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     6121 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     6181 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     6241 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catg
//
