LOCUS       sequence-272323-        7093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid was constructed to study cortactin interactions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7093)
  AUTHORS   Weed SA, Karginov AV, Schafer DA, Weaver AM, Kinley AW, Cooper JA, 
            Parsons JT
  TITLE     Cortactin localization to sites of actin assembly in lamellipodia 
            requires interactions with F-actin and the Arp2/3 complex.
  JOURNAL   J Cell Biol. 2000 Oct 2;151(1):29-40.
  PUBMED    11018051
REFERENCE   2  (bases 1 to 7093)
  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 
            Biol."; date: "2000-10-2"; volume: "151(1)"; pages: "29-40"
COMMENT     Sequence Label: pcDNA3-Flag-Cortactin
FEATURES             Location/Qualifiers
     source          1..7093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      894..903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             903..926
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2646..2664)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2647..2664)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2684..2701)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2690..2914
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2960..3388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3047..3066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3257..3278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3397..3417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3402..3731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3582..3717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3644..3663
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3798..4592
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3852..3871)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4462..4481
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4766..4887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4803..4822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4857..4876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4936..4952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4936..4952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4949..4971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4960..4976
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4984..4990,4991..5008,5009..5014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5029..5050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5167..5184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5338..5926)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5418..5437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6097..6888,6889..6957))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6720..6739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6958..7062)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctgccacca
      901 tggactacaa ggacgacgat gacaagggac ctaagcttgg taccatgtgg aaagcctctg
      961 caggccatgc tgtgtccatc acgcaggatg atggaggagc tgatgactgg gagactgatc
     1021 ctgattttgt gaatgatgtg agtgaaaagg agcagagatg gggtgctaaa accgtgcagg
     1081 gatcggggca ccaggaacac atcaacattc acaagcttcg agagaatgtc ttccaagaac
     1141 accagacgct caaggagaag gagctggaaa cgggacccaa ggcttcccac ggctatggcg
     1201 ggaagttcgg tgtggagcag gataggatgg acagatcagc cgtgggccat gagtaccagt
     1261 cgaagctttc caagcactgc tcacaagtgg actcggtccg gggcttcgga ggcaagttcg
     1321 gtgtccagat ggacagggtg gatcagtctg ctgtaggctt tgaataccag gggaagactg
     1381 agaagcatgc ctcccagaaa gactactcta gtggcttcgg tggcaaatac ggtgtgcaag
     1441 ctgaccgtgt agacaagagt gccgtgggct ttgactacca gggcaagacg gagaagcatg
     1501 agtctcagaa agattactcc aaaggttttg gtggcaaata tgggattgac aaggacaagg
     1561 tggataaaag tgctgtgggc tttgagtatc aaggcaagac agagaagcac gaatcccaga
     1621 aagactatgt aaaaggcttt ggaggaaagt ttggtgtgca gacagacaga caggacaagt
     1681 gtgcccttgg ctgggaccat caggagaagc tgcagctgca tgaatcccaa aaagactata
     1741 agactggttt cggaggcaaa tttggtgttc agtccgagag gcaggactcc tccgctgtgg
     1801 ggtttgatta caaggagaga ttggccaagc acgagcccca gcaagactat gccaaaggat
     1861 tcggcgggaa gtatggggtg cagaaggatc ggatggacaa gaatgcatcc acctttgaag
     1921 aagtggtcca ggtgccatct gcctatcaga agactgtccc cattgaggcc gtaaccagca
     1981 aaaccagtaa tatccgtgct aactttgaaa acctggcaaa ggagagagag caggaggaca
     2041 ggcggaaggc agaagccgag agagctcagc ggatggccaa agaaagacag gagcaggagg
     2101 aggcgcgcag gaagctggaa gagcaagcca gagccaagaa gcagacgccc cctgcatccc
     2161 ctagtcctca accaattgaa gacagaccac cctccagccc catctatgag gatgcagctc
     2221 cgttcaaggc cgagccgagc taccgaggta gcgaacctga gcctgagtac agcatcgagg
     2281 ccgcaggcat tcctgaggct ggcagccagc aaggcctgac ctatacatca gagcccgtgt
     2341 acgagactac agaggctcct ggccactatc aagcagagga tgacacctac gatgggtatg
     2401 agagtgacct gggcatcaca gccatcgccc tgtatgacta ccaggctgct ggcgatgatg
     2461 agatctcctt tgaccctgat gacatcatca ccaacataga aatgattgac gatggctggt
     2521 ggcgtggggt gtgcaagggc agatacgggc tcttcccagc caactatgtg gagctgcggc
     2581 agtaggaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
     2641 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     2701 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2761 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2821 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2881 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2941 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3001 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3061 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3121 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3181 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3241 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3301 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3361 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3481 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3601 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3661 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3721 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3781 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3841 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3901 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3961 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4021 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4081 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4141 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4201 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4261 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4321 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4381 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4441 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4501 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4561 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4621 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4681 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4741 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4801 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4861 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4921 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4981 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5041 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5101 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5161 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5221 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6241 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7081 gccacctgac gtc
//
