LOCUS       sequence-138522-        8733 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 8733)
  AUTHORS   Jacob KK, Whittaker J, Stanley FM
  TITLE     Insulin receptor tyrosine kinase activity and phosphorylation of 
            tyrosines 1162 and 1163 are required for insulin-increased prolactin
            gene expression.
  JOURNAL   Mol Cell Endocrinol. 2002 Jan 15. 186(1):7-16.
  PUBMED    11850117
REFERENCE   2  (bases 1 to 8733)
  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: "Mol Cell 
            Endocrinol. 2002 Jan 15. 186(1):7-16."
COMMENT     Sequence Label: HIR WT
FEATURES             Location/Qualifiers
     source          1..8733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2026..2044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2082..2104)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2204..2223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2388..2767
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2768..2971
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2922..2942
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3016..3035
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3016..3034
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          7344..7409
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7539..7559
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7831..7965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7868..7887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7922..7941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8203..8338
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2041 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2101 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2161 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2221 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2281 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2341 gcctcttcgc tattacgcca gcgatgtacg ggccagatat acgcgttgac attgattatt
     2401 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     2461 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     2521 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     2581 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     2641 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2701 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2761 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     2821 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     2881 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     2941 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     3001 ctggcttatc gaaattaata cgactcacta tagggagacc caagcttcct taaggacatg
     3061 ggcaccgggg gccggcgggg ggcggcggcc gcgccgctgc tggtggcggt ggccgcgctg
     3121 ctactgggcg ccgcgggcca cctgtacccc ggagaggtgt gtcccggcat ggatatccgg
     3181 aacaacctca ctaggttgca tgagctggag aattgctctg tcatcgaagg acacttgcag
     3241 atactcttga tgttcaaaac gaggcccgaa gatttccgag acctcagttt ccccaaactc
     3301 atcatgatca ctgattactt gctgctcttc cgggtctatg ggctcgagag cctgaaggac
     3361 ctgttcccca acctcacggt catccgggga tcacgactgt tctttaacta cgcgctggtc
     3421 atcttcgaga tggttcacct caaggaactc ggcctctaca acctgatgaa catcacccgg
     3481 ggttctgtcc gcatcgagaa gaacaatgag ctctgttact tggccactat cgactggtcc
     3541 cgtatcctgg attccgtgga ggataattac atcgtgttga acaaagatga caacgaggag
     3601 tgtggagaca tctgtccggg taccgcgaag ggcaagacca actgccccgc caccgtcatc
     3661 aacgggcagt ttgtcgaacg atgttggact catagtcact gccagaaagt ttgcccgacc
     3721 atctgtaagt cacacggctg caccgccgaa ggcctctgtt gccacagcga gtgcctgggc
     3781 aactgttctc agcccgacga ccccaccaag tgcgtggcct gccgcaactt ctacctggac
     3841 ggcaggtgtg tggagacctg cccgcccccg tactaccact tccaggactg gcgctgtgtg
     3901 aacttcagct tctgccagga cctgcaccac aaatgcaaga actcgcggag gcagggctgc
     3961 caccagtacg tcattcacaa caacaagtgc atccctgagt gtccctccgg gtacacgatg
     4021 aattccagca acttgctgtg caccccatgc ctgggtccct gtcccaaggt gtgccacctc
     4081 ctagaaggcg agaagaccat cgactcggtg acgtctgccc aggagctccg aggatgcacc
     4141 gtcatcaacg ggagtctgat catcaacatt cgaggaggca acaatctggc agctgagcta
     4201 gaagccaacc tcggcctcat tgaagaaatt tcagggtatc taaaaatccg ccgatcctac
     4261 gctctggtgt cactttcctt cttccggaag ttacgtctga ttcgaggaga gaccttggaa
     4321 attgggaact actccttcta tgccttggac aaccagaacc taaggcagct ctgggactgg
     4381 agcaaacaca acctcaccat cactcagggg aaactcttct tccactataa ccccaaactc
     4441 tgcttgtcag aaatccacaa gatggaagaa gtttcaggaa ccaaggggcg ccaggagaga
     4501 aacgacattg ccctgaagac caatggggac caggcatcct gtgaaaatga gttacttaaa
     4561 ttttcttaca ttcggacatc ttttgacaag atcttgctga gatgggagcc gtactggccc
     4621 cccgacttcc gagacctctt ggggttcatg ctgttctaca aagaggcccc ttatcagaat
     4681 gtgacggagt tcgacgggca ggatgcgtgt ggttccaaca gttggacggt ggtagacatt
     4741 gacccacccc tgaggtccaa cgaccccaaa tcacagaacc acccagggtg gctgatgcgg
     4801 ggtctcaagc cctggaccca gtatgccatc tttgtgaaga ccctggtcac cttttcggat
     4861 gaacgccgga cctatggggc caagagtgac atcatttatg tccagacaga tgccaccaac
     4921 ccctctgtgc ccctggatcc aatctcagtg tctaactcat catcccagat tattctgaag
     4981 tggaaaccac cctccgaccc caatggcaac atcacccact acctggtttt ctgggagagg
     5041 caggcggaag acagtgagct gttcgagctg gattattgcc tcaaagggct gaagctgccc
     5101 tcgaggacct ggtctccacc attcgagtct gaagattctc agaagcacaa ccagagtgag
     5161 tatgaggatt cggccggcga atgctgctcc tgtccaaaga cagactctca gatcctgaag
     5221 gagctggagg agtcctcgtt taggaagacg tttgaggatt acctgcacaa cgtggttttc
     5281 gtccccagaa aaacctcttc aggcactggt gccgaggacc ctaggccatc tcggaaacgc
     5341 aggtcccttg gcgatgttgg gaatgtgacg gtggccgtgc ccacggtggc agctttcccc
     5401 aacacttcct cgaccagcgt gcccacgagt ccggaggagc acaggccttt tgagaaggtg
     5461 gtgaacaagg agtcgctggt catctccggc ttgcgacact tcacgggcta tcgcatcgag
     5521 ctgcaggctt gcaaccagga cacccctgag gaacggtgca gtgtggcagc ctacgtcagt
     5581 gcgaggacca tgcctgaagc caaggctgat gacattgttg gccctgtgac gcatgaaatc
     5641 tttgagaaca acgtcgtcca cttgatgtgg caggagccga aggagcccaa tggtctgatc
     5701 gtgctgtatg aagtgagtta tcggcgatat ggtgatgagg agctgcatct ctgcgtctcc
     5761 cgcaagcact tcgctctgga acggggctgc aggctgcgtg ggctgtcacc ggggaactac
     5821 agcgtgcgaa tccgggccac ctcccttgcg ggcaacggct cttggacgga acccacctat
     5881 ttctacgtga cagactattt agacgtcccg tcaaatattg caaaaattat catcggcccc
     5941 ctcatctttg tctttctctt cagtgttgtg attggaagta tttatctatt cctgagaaag
     6001 aggcagccag atgggccgct gggaccgctt tacgcttctt caaaccctga gtatctcagt
     6061 gccagtgatg tgtttccatg ctctgtgtac gtgccggacg agtgggaggt gtctcgagag
     6121 aagatcaccc tccttcgaga gctggggcag ggctccttcg gcatggtgta tgagggcaat
     6181 gccagggaca tcatcaaggg tgaggcagag acccgcgtgg cggtgaagac ggtcaacgag
     6241 tcagccagtc tccgagagcg gattgagttc ctcaatgagg cctcggtcat gaagggcttc
     6301 acctgccatc acgtggtgcg cctcctggga gtggtgtcca agggccagcc cacgctggtg
     6361 gtgatggagc tgatggctca cggagacctg aagagctacc tccgttctct gcggccagag
     6421 gctgagaata atcctggccg ccctccccct acccttcaag agatgattca gatggcggca
     6481 gagattgctg acgggatggc ctacctgaac gccaagaagt ttgtgcatcg ggacctggca
     6541 gcgagaaact gcatggtcgc ccatgatttt actgtcaaaa ttggagactt tggaatgacc
     6601 agagacatct atgaaacgga ttactaccgg aaagggggca agggtctgct ccctgtacgg
     6661 tggatggcac cggagtccct gaaggatggg gtcttcacca cttcttctga catgtggtcc
     6721 tttggcgtgg tcctttggga aatcaccagc ttggcagaac agccttacca aggcctgtct
     6781 aatgaacagg tgttgaaatt tgtcatggat ggagggtatc tggatcaacc cgacaactgt
     6841 ccagagagag tcactgacct catgcgcatg tgctggcaat tcaaccccaa gatgaggcca
     6901 accttcctgg agattgtcaa cctgctcaag gacgacctgc accccagctt tccagaggtg
     6961 tcgttcttcc acagcgagga gaacaaggct cccgagagtg aggagctgga gatggagttt
     7021 gaggacatgg agaatgtgcc cctggaccgt tcctcgcact gtcagaggga ggaggcgggg
     7081 ggccgggatg gagggtcctc gctgggtttc aagcggagct acgaggaaca catcccttac
     7141 acacacatga acggaggcaa gaaaaacggg cggattctga ccttgcctcg gtccaatcct
     7201 tcctaagtcg agggatcttc catacctacc agttctgcgc ctgcaggtcg cggccgcgac
     7261 tctagaggat ctttgtgaag gaaccttact tctgtggtgt gacataattg gacaaactac
     7321 ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac
     7381 tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc
     7441 agtggtggaa tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg
     7501 atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag
     7561 aagaccccaa ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta
     7621 atagaactct tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca
     7681 agaaaattat ggaaaaatat ttgatgtata gtgccttgac tagagatcat aatcagccat
     7741 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     7801 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     7861 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7921 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcccgcc atggtatcaa
     7981 cgccatattt ctatttacag tagggacctc ttcgttgtgt agcctggcct ccgcttactc
     8041 tggagaaaaa gaagagaggc attgtagagg cttccagagg caacttgtca aaacaggact
     8101 gcttctattt ctgtcacact gtctggccct gtcacaaggt ccagcacctc cataccccct
     8161 ttaataagca gtttgggaac gggtgcgggt cttactccgc ccatcccgcc cctaactccg
     8221 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     8281 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     8341 ggcttttgca aaaagctaat tcggcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     8521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctgaagccag ttaccagtgg
     8581 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagagat agttaccgga
     8641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcact gcattaatga atcggccaac
     8701 gcgcggggag aggcggtttg cgtattgggc gct
//
