LOCUS       sequence-350969-        7624 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 7624)
  AUTHORS   Araki E, Sun XJ, Haag BL, Chuang LM, Zhang Y, Yang-Feng TL, White 
            MF, Kahn CR
  TITLE     Human skeletal muscle insulin receptor substrate-1. Characterization
            of the cDNA, gene, and chromosomal localization.
  JOURNAL   Diabetes. 1993 Jul . 42(7):1041-54.
  PUBMED    8513971
REFERENCE   2  (bases 1 to 7624)
  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: "Diabetes. 
            1993 Jul . 42(7):1041-54."
COMMENT     Sequence Label: pBS human IRS-1
FEATURES             Location/Qualifiers
     source          1..7624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..146
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     138..155
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     139..155
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     165..184
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        165..183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     216..232
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     216..232
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     regulatory      4358..4367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4929..4945)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4930..4946)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4975..4995)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4975..4993)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5014..5030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5014..5030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5027..5049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5038..5054
                     /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(5062..5068,5069..5086,5087..5092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5107..5128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5245..5262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5416..6004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5496..5515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6175..6966,6967..7035))
                     /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     6798..6817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7036..7140)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7167..7622
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7254..7273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7464..7485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
       61 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      121 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
      181 agggcgaatt ggagctccac cgcggtggcg gccgctctag aactagtgga tcccccgggc
      241 tgcaggaatt cccggcggcg cggtcggagg gggccggcgc gcagagccag acgccgccgc
      301 ttgttttggt tggggctctc ggcaactctc cgaggaggag gaggaggagg gaggagggga
      361 gaagtaactg cagcggcagc gcctcccgag gaacaggcgt cttccccgaa cccttcccaa
      421 acctccccca tcccctctcg cccttgtccc ctcccctcct ccccagccgc ctggagcgag
      481 gggcagggat gagtctgtcc ctccggccgg tccccagctg cagtggctgc ccggtatcgt
      541 ttcgcatgga aaagccactt tctccacccg ccgagatggg cccggatggg ggctgcagag
      601 gacgcgcccg cgggcggcgg cagcagcagc agcagcagca gcagcaacag caacagccgc
      661 agcgccgcgg tctctgcgac tgagctggta tttgggcggc tggtggcggc tgggacggtt
      721 ggggggtggg aggaggcgaa ggaggaggga gaaccccgtg caacgttggg acttggcaac
      781 ccgcctcccc ctgcccaagg atatttaatt tgcctcggga atcgctgctt ccagagggga
      841 actcaggagg gaaggcgcgc gcgcgcgcgc gctcctggag gggcaccgca gggacccccg
      901 actgtcgcct ccctgtgccg gactccagcc ggggcgacga gagatgcatc ttcgctcctt
      961 cctggtggcg gcggcggctg agaggagact tggctctcgg aggatcgggg ctgccctcac
     1021 cccggacgca ctgcctcccc gccgggcgtg aagcgcccga aaactccggt cgggctctct
     1081 cctgggctca gcagctgcgt cctccttcag ctgcccctcc ccggcgcggg gggcggcgtg
     1141 gatttcagag tcggggtttc tgctgcctcc agccctgttt gcatgtgccg ggccgcggcg
     1201 aggagcctcc gccccccacc cggttgtttt tcggagcctc cctctgctca gcgttggtgg
     1261 tggcggtggc agcatggcga gccctccgga gagcgatggc ttctcggacg tgcgcaaggt
     1321 gggctacctg cgcaaaccca agagcatgca caaacgcttc ttcgtactgc gcgcggccag
     1381 cgaggctggg ggcccggcgc gcctcgagta ctacgagaac gagaagaagt ggcggcacaa
     1441 gtcgagcgcc cccaaacgct cgatccccct tgagagctgc ttcaacatca acaagcgggc
     1501 tgactccaag aacaagcacc tggtggctct ctacacccgg gacgagcact ttgccatcgc
     1561 ggcggacagc gaggccgagc aagacagctg gtaccaggct ctcctacagc tgcacaaccg
     1621 tgctaagggc caccacgacg gagctgcggc cctcggggcg ggaggtggtg ggggcagctg
     1681 cagcggcagc tccggccttg gtgaggctgg ggaggacttg agctacggtg acgtgccccc
     1741 aggacccgca ttcaaagagg tctggcaagt gatcctgaag cccaagggcc tgggtcagac
     1801 aaagaacctg attggtatct accgcctttg cctgaccagc aagaccatca gcttcgtgaa
     1861 gctgaactcg gaggcagcgg ccgtggtgct gcagctgatg aacatcaggc gctgtggcca
     1921 ctcggaaaac ttcttcttca tcgaggtggg ccgttctgcc gtgacggggc ccggggagtt
     1981 ctggatgcag gtggatgact ctgtggtggc ccagaacatg cacgagacca tcctggaggc
     2041 catgcgggcc atgagtgatg agttccgccc tcgcagcaag agccagtcct cgtccaactg
     2101 ctctaacccc atcagcgtcc ccctgcgccg gcaccatctc aacaatcccc cgcccagcca
     2161 ggtggggctg acccgccgat cacgcactga gagcatcacc gccacctccc cggccagcat
     2221 ggtgggcggg aagccaggct ccttccgtgt ccgcgcctcc agtgacggcg aaggcaccat
     2281 gtcccgccca gcctcggtgg acggcagccc tgtgagtccc agcaccaaca gaacccacgc
     2341 ccaccggcat cggggcagcg cccggctgca ccccccgctc aaccacagcc gctccatccc
     2401 catgccggct tcccgctgct cgccttcggc caccagcccg gtcagtctgt cgtccagtag
     2461 caccagtggc catggctcca cctcggattg tctcttccca cggcgatcta gtgcttcggt
     2521 gtctggttcc cccagcgatg gcggtttcat ctcctcggat gagtatggct ccagtccctg
     2581 cgatttccgg agttccttcc gcagtgtcac tccggattcc ctgggccaca ccccaccagc
     2641 ccgcggtgag gaggagctaa gcaactatat ctgcatgggt ggcaaggggc cctccaccct
     2701 gaccgccccc aacggtcact acattttgtc tcggggtggc aatggccacc gctgcacccc
     2761 aggaacaggc ttgggcacga gtccagcctt ggctggggat gaagcagcca gtgctgcaga
     2821 tctggataat cggttccgaa agagaactca ctcggcaggc acatccccta ccattaccca
     2881 ccagaagacc ccgtcccagt cctcagtggc ttccattgag gagtacacag agatgatgcc
     2941 tgcctaccca ccaggaggtg gcagtggagg ccgactgccg ggacacaggc actccgcctt
     3001 cgtgcccacc cgctcctacc cagaggaggg tctggaaatg caccccttgg agcgtcgggg
     3061 ggggcaccac cgcccagaca gctccaccct ccacacggat gatggctaca tgcccatgtc
     3121 cccaggggtg gccccagtgc ccagtggccg aaagggcagt ggagactata tgcccatgag
     3181 ccccaagagc gtatctgccc cacagcagat catcaatccc atcagacgcc atccccagag
     3241 agtggacccc aatggctaca tgatgatgtc ccccagcggt ggctgctctc ctgacattgg
     3301 aggtggcccc agcagcagca gcagcagcag caacgccgtc ccttccggga ccagctatgg
     3361 aaagctgtgg acaaacgggg tagggggcca ccactctcat gtcttgcctc accccaaacc
     3421 cccagtggag agcagcggtg gtaagctctt accttgcaca ggtgactaca tgaacatgtc
     3481 accagtgggg gactccaaca ccagcagccc ctccgactgc tactacggcc ctgaggaccc
     3541 ccagcacaag ccagtcctct cctactactc attgccaaga tcctttaagc acacccagcg
     3601 ccccggggag ccggaggagg gtgcccggca tcagcacctc cgcctttcca ctagctctgg
     3661 tcgccttctc tatgctgcaa cagcagatga ttcttcctct tccaccagca gcgacagcct
     3721 gggtggggga tactgcgggg ctaggctgga gcccagcctt ccacatcccc accatcaggt
     3781 tctgcagccc catctgcctc gaaaggtgga cacagctgct cagaccaata gccgcctggc
     3841 ccggcccacg aggctgtccc tgggggatcc caaggccagc accttacctc gggcccgaga
     3901 gcagcagcag cagcagcagc ccttgctgca ccctccagag cccaagagcc cgggggaata
     3961 tgtcaatatt gaatttggga gtgatcagtc tggctacttg tctggcccgg tggctttcca
     4021 cagctcacct tctgtcaggt gtccatccca gctccagcca gctcccagag aggaagagac
     4081 tggcactgag gagtacatga agatggacct ggggccgggc cggagggcag cctggcagga
     4141 gagcactggg gtcgagatgg gcagactggg ccctgcacct cccggggctg ctagcatttg
     4201 caggcctacc cgggcagtgc ccagcagccg gggtgactac atgaccatgc agatgagttg
     4261 tccccgtcag agctacgtgg acacctcgcc agctgcccct gtaagctatg ctgacatgcg
     4321 aacaggcatt gctgcagagg aggtgagcct gcccagggcc accatggctg ctgcctcctc
     4381 atcctcagca gcctctgctt ccccgactgg gcctcaaggg gcagcagagc tggctgccca
     4441 ctcgtccctg ctggggggcc cacaaggacc tgggggcatg agcgccttca cccgggtgaa
     4501 cctcagtcct aaccgcaacc agagtgccaa agtgatccgt gcagacccac aagggtgccg
     4561 gcggaggcat agctccgaga ctttctcctc aacacccagt gccacccggg tgggcaacac
     4621 agtgcccttt ggagcggggg cagcagtagg gggcggtggc ggtagcagca gcagcagcga
     4681 ggatgtgaaa cgccacagct ctgcttcctt tgagaatgtg tggctgaggc ctggggagct
     4741 tgggggagcc cccaaggagc cagccaaact gtgtggggct gctgggggtt tggagaatgg
     4801 tcttaactac atagacctgg atttggtcaa ggacttcaaa cagtgccctc aggagtgcac
     4861 ccctgaaccg cagcctcccc cacccccacc ccctcatcaa ccccaaagga attcgatatc
     4921 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccagctttt gttcccttta
     4981 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5041 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5221 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7141 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     7201 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     7261 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     7321 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     7381 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     7441 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     7501 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     7561 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     7621 ttcc
//
