LOCUS       sequence-174150-        6352 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 6352)
  AUTHORS   Mariotti A, Kedeshian PA, Dans M, Curatola AM, Gagnoux-Palacios L, 
            Giancotti FG
  TITLE     EGF-R signaling through Fyn kinase disrupts the function of integrin
            alpha6beta4 at hemidesmosomes: role in epithelial cell migration and
            carcinoma invasion.
  JOURNAL   J Cell Biol. 2001 Oct 29. 155(3):447-58.
  PUBMED    11684709
REFERENCE   2  (bases 1 to 6352)
  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. 2001 Oct 29. 155(3):447-58."
COMMENT     Sequence Label: pRK5 c-Fyn
FEATURES             Location/Qualifiers
     source          1..6352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..90
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    207..228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(243..248,249..266,267..273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    281..297
                     /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)."
     primer_bind     286..308
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     305..321
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     305..321
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        350..729
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        730..933
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     884..904
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     930..954
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1164..1182
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1164..1181
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1195..1214
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2988..2997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3001..3135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3038..3057)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3092..3111
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3204..3561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3204..3395
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3227..3247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3412..3547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3581..3598)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3581..3597)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3590..3612)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3810..4265
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3897..3916)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4107..4128
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4282..4301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4401..4423
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4461..4479)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4547..4651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4652..4720,4721..5512)
                     /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     complement(4870..4889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5683..6271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6172..6191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
       61 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
      121 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      181 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      241 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      301 cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg acattgatta
      361 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      421 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      481 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      541 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      601 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      661 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      721 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      781 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      841 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      901 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      961 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
     1021 ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc
     1081 tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa ttaatacata
     1141 accttatgta tcatacacat acgatttagg tgacactata gaataacatc cactttgcct
     1201 ttctctccac aggtgtccac tcccaggtcc aactgcacct cggttctatc gcgattgaat
     1261 tccccgggga tcctctagcg cgtgacggat cgttggcgga aattttggaa tttagaccat
     1321 gggctgtgtg caatgtaagg ataaagaagc aacaaaactg acggaggaga gggacggcag
     1381 cctgaaccag agctctgggt accgctatgg cacagacccc acccctcagc actaccccag
     1441 cttcggtgtg acctccatcc ccaactacaa caacttccac gcagccgggg gccaaggact
     1501 caccgtcttt ggaggtgtga actcttcgtc tcatacgggg accttgcgta cgagaggagg
     1561 aacaggagtg acactctttg tggcccttta tgactatgaa gcacggacag aagatgacct
     1621 gagttttcac aaaggagaaa aatttcaaat attgaacagc tcggaaggag attggtggga
     1681 agcccgctcc ttgacaactg gagagacagg ttacattccc agcaattatg tggctccagt
     1741 tgactctatc caggcagaag agtggtactt tggaaaactt ggccgaaaag atgctgagcg
     1801 acagctattg tcctttggaa acccaagagg tacctttctt atccgcgaga gtgaaaccac
     1861 caaaggtgcc tattcacttt ctatccgtga ttgggatgat atgaaaggag accatgtcaa
     1921 acattataaa attcgcaaac ttgacaatgg tggatactac attaccaccc gggcccagtt
     1981 tgaaacactt cagcagcttg tacaacatta ctcagagaga gctgcaggtc tctgctgccg
     2041 cctagtagtt ccctgtcaca aagggatgcc aaggcttacc gatctgtctg tcaaaaccaa
     2101 agatgtctgg gaaatccctc gagaatccct gcagttgatc aagagactgg gaaatgggca
     2161 gtttggggaa gtatggatgg gtacctggaa tggaaacaca aaagtagcca taaagactct
     2221 taaaccaggc acaatgtccc ccgaatcatt ccttgaggaa gcgcagatca tgaagaagct
     2281 gaagcacgac aagctggtcc agctctatgc agtggtgtct gaggagccca tctacatcgt
     2341 caccgagtat atgaacaaag gaagtttact ggatttctta aaagatggag aaggaagagc
     2401 tctgaaatta ccaaatcttg tggacatggc agcacaggtg gctgcaggaa tggcttacat
     2461 cgagcgcatg aattatatcc atagagatct gcgatcagca aacattctag tggggaatgg
     2521 actcatatgc aagattgctg acttcggatt ggcccgattg atagaagaca atgagtacac
     2581 agcaagacaa ggtgcaaagt tccccatcaa gtggacggcc cccgaggcag ccctgtacgg
     2641 gaggttcaca atcaagtctg acgtgtggtc ttttggaatc ttactcacag agctggtcac
     2701 caaaggaaga gtgccatacc caggcatgaa caaccgggag gtgctggagc aggtggagcg
     2761 aggctacagg atgccctgcc cgcaggactg ccccatctct ctgcatgagc tcatgatcca
     2821 ctgctggaaa aaggaccctg aagaacgccc cacttttgag tacttgcaga gcttcctgga
     2881 agactacttt accgcgacag agccccagta ccaacctggt gaaaacctgt aaggcccggg
     2941 tctgcggaga gagggacgcg ctagagtcga cctgcagccc aagcttggcc gccatggccc
     3001 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3061 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3121 tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat aacctctgaa
     3181 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3241 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3481 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3541 ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc gttttacaac
     3601 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     3661 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3721 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3781 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3841 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3901 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3961 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4021 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4081 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4141 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4201 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4261 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4321 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4381 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4441 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4501 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4561 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4621 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4681 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4741 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4801 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4861 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4921 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4981 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5041 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5101 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5161 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5221 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5281 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5341 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5401 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5461 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5521 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5701 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5761 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5821 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5881 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5941 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6001 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6061 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6121 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6181 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6241 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6301 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ct
//
