LOCUS       sequence-175334-        8257 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 8257)
  AUTHORS   Dans M, Gagnoux-Palacios L, Blaikie P, Klein S, Mariotti A, 
            Giancotti FG
  TITLE     Tyrosine phosphorylation of the beta 4 integrin cytoplasmic domain 
            mediates Shc signaling to extracellular signal-regulated kinase and 
            antagonizes formation of hemidesmosomes.
  JOURNAL   J Biol Chem. 2001 Jan 12. 276(2):1494-502.
  PUBMED    11044453
REFERENCE   2  (bases 1 to 8257)
  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 Biol 
            Chem. 2001 Jan 12. 276(2):1494-502."
COMMENT     Sequence Label: pRK5 alpha6
FEATURES             Location/Qualifiers
     source          1..8257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..89
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    206..227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(242..247,248..265,266..272)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    280..296
                     /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     285..307
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     304..320
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     304..320
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        349..728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        729..932
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     883..903
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     929..953
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1163..1181
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1163..1180
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1194..1213
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4892..4901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4905..5039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4942..4961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4996..5015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5108..5465
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5108..5299
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5131..5151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5316..5451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5485..5502)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5485..5501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5494..5516)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5714..6169
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5801..5820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6011..6032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6186..6205)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6305..6327
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6365..6383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6451..6555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6556..6624,6625..7416)
                     /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(6774..6793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7587..8175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8076..8095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
       61 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      121 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      181 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      241 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      301 acacaggaaa cagctatgac atgattacga attaattcga gctcgcccga cattgattat
      361 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      421 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      481 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      541 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      601 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      661 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      721 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      781 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      841 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      901 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
      961 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg
     1021 gccgggaacg gtgcattgga acgcggattc cccgtgccaa gagtgacgta agtaccgcct
     1081 atagagtcta taggcccacc cccttggctt cgttagaacg cggctacaat taatacataa
     1141 ccttatgtat catacacata cgatttaggt gacactatag aataacatcc actttgcctt
     1201 tctctccaca ggtgtccact cccaggtcca actgcacctc ggttctatcg attgaattcc
     1261 ccggggatcc tctagagtcg agatccattg tgctctaaag gccggacgga gagcgcgacc
     1321 cgtcccgggg gtggggccgg gcgcagcggc gagaggaggc gaaggtggct gcggtagcag
     1381 cagcgcggca gcctcggacc cagcccggag cgcagggcgg ccgctgcagg tccccgctcc
     1441 cctccccgtg cgtccgccca tggccgccgc cgggcagctg tgcttgctct acctgtcggc
     1501 ggggctcctg tcccggctcg gcgcagcctt caacttggac actcgggagg acaacgtgat
     1561 ccggaaatat ggagaccccg ggagcctctt cggcttctcg ctggccatgc actggcaact
     1621 gcagcccgag gacaagcggc tgttgctcgt gggggccccg cgggcagaag cgcttccact
     1681 gcagagagcc aacagaacgg gagggctgta cagctgcgac atcaccgccc gggggccatg
     1741 cacgcggatc gagtttgata acgatgctga ccccacgtca gaaagcaagg aagatcagtg
     1801 gatgggggtc accgtccaga gccaaggtcc agggggcaag gtcgtgacat gtgctcaccg
     1861 atatgaaaaa aggcagcatg ttaatacgaa gcaggaatcc cgagacatct ttgggcggtg
     1921 ttatgtcctg agtcagaatc tcaggattga agacgatatg gatgggggag attggagctt
     1981 ttgtgatggg cgattgagag gccatgagaa atttggctct tgccagcaag gtgtagcagc
     2041 tacttttact aaagactttc attacattgt atttggagcc ccgggtactt ataactggaa
     2101 agggattgtt cgtgtagagc aaaagaataa cacttttttt gacatgaaca tctttgaaga
     2161 tgggccttat gaagttggtg gagagactga gcatgatgaa agtctcgttc ctgttcctgc
     2221 taacagttac ttaggttttt ctttggactc agggaaaggt attgtttcta aagatgagat
     2281 cacttttgta tctggtgctc ccagagccaa tcacagtgga gccgtggttt tgctgaagag
     2341 agacatgaag tctgcacatc tcctccctga gcacatattc gatggagaag gtctggcctc
     2401 ttcatttggc tatgatgtgg cggtggtgga cctcaacaag gatgggtggc aagatatagt
     2461 tattggagcc ccacagtatt ttgatagaga tggagaagtt ggaggtgcag tgtatgtcta
     2521 catgaaccag caaggcagat ggaataatgt gaagccaatt cgtcttaatg gaaccaaaga
     2581 ttctatgttt ggcattgcag taaaaaatat tggagatatt aatcaagatg gctacccaga
     2641 tattgcagtt ggagctccgt atgatgactt gggaaaggtt tttatctatc atggatctgc
     2701 aaatggaata aataccaaac caacacaggt tctcaagggt atatcacctt attttggata
     2761 ttcaattgct ggaaacatgg accttgatcg aaattcctac cctgatgttg ctgttggttc
     2821 cctctcagat tcagtaacta ttttcagatc ccggcctgtg attaatattc agaaaaccat
     2881 cacagtaact cctaacagaa ttgacctccg ccagaaaaca gcgtgtgggg cgcctagtgg
     2941 gatatgcctc caggttaaat cctgttttga atatactgct aaccccgctg gttataatcc
     3001 ttcaatatca attgtgggca cacttgaagc tgaaaaagaa agaagaaaat ctgggctatc
     3061 ctcaagagtt cagtttcgaa accaaggttc tgagcccaaa tatactcaag aactaactct
     3121 gaagaggcag aaacagaaag tgtgcatgga ggaaaccctg tggctacagg ataatatcag
     3181 agataaactg cgtcccattc ccataactgc ctcagtggag atccaagagc caagctctcg
     3241 taggcgagtg aattcacttc cagaagttct tccaattctg aattcagatg aacccaagac
     3301 agctcatatt gatgttcact tcttaaaaga gggatgtgga gacgacaatg tatgtaacag
     3361 caaccttaaa ctagaatata aattttgcac ccgagaagga aatcaagaca aattttctta
     3421 tttaccaatt caaaaaggtg taccagaact agttctaaaa gatcagaagg atattgcttt
     3481 agaaataaca gtgacaaaca gcccttccaa cccaaggaat cccacaaaag atggcgatga
     3541 cgcccatgag gctaaactga ttgcaacgtt tccagacact ttaacctatt ctgcatatag
     3601 agaactgagg gctttccctg agaaacagtt gagttgtgtt gccaaccaga atggctcgca
     3661 agctgactgt gagctcggaa atccttttaa aagaaattca aatgtcactt tttatttggt
     3721 tttaagtaca actgaagtca cctttgacac cccatatctg gatattaatc tgaagttaga
     3781 aacaacaagc aatcaagata atttggctcc aattacagct aaagcaaaag tggttattga
     3841 actgctttta tcggtctcgg gagttgctaa accttcccag gtgtattttg gaggtacagt
     3901 tgttggcgag caagctatga aatctgaaga tgaagtggga agtttaatag agtatgaatt
     3961 cagggtaata aacttaggta aacctcttac aaacctcggc acagcaacct tgaacattca
     4021 gtggccaaaa gaaattagca atgggaaatg gttgctttat ttggtgaaag tagaatccaa
     4081 aggattggaa aaggtaactt gtgagccaca aaaggagata aactccctga acctaacgga
     4141 gtctcacaac tcaagaaaga aacgggaaat tactgaaaaa cagatagatg ataacagaaa
     4201 attttcttta tttgctgaaa gaaaatacca gactcttaac tgtagcgtga acgtgaactg
     4261 tgtgaacatc agatgcccgc tgcgggggct ggacagcaag gcgtctctta ttttgcgctc
     4321 gaggttatgg aacagcacat ttctagagga atattccaaa ctgaactact tggacattct
     4381 catgcgagcc ttcattgatg tgactgctgc tgccgaaaat atcaggctgc caaatgcagg
     4441 cactcaggtt cgagtgactg tgtttccctc aaagactgta gctcagtatt cgggagtacc
     4501 ttggtggatc atcctagtgg ctattctcgc tgggatcttg atgcttgctt tattagtgtt
     4561 tatactatgg aagtgtggtt tcttcaagag aaataagaaa gatcattatg atgccacata
     4621 tcacaaggct gagatccatg ctcagccatc tgataaagag aggcttactt ctgatgcata
     4681 gtattgatct acttctgtaa ttgtgtggat tctttaaacg ctctaggtac gatgacagtg
     4741 ttccccgata ccatgctgta aggatccgga aagaagagcg agagatcaaa gatgaaaagt
     4801 atattgataa ccttgaaaaa aaacagtgga tcacaaagtg gaacgaaaat gaaagctact
     4861 catagcgggg gcctaaaaaa aaaaaagctt ggccgccatg gcccaacttg tttattgcag
     4921 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4981 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     5041 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     5101 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5161 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5281 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5341 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5401 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5461 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5521 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5581 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5641 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5701 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     6061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6181 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6241 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6301 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6361 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6421 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6481 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6541 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6601 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6661 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6721 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6781 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6841 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6901 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6961 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7021 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7081 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7141 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7201 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7261 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7321 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7381 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7441 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7501 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7561 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7621 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7681 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7741 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7801 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7861 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7921 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7981 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8041 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8101 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8161 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8221 ccttttgctc acatgttctt tcctgcgtta tcccctg
//
