LOCUS       sequence-200837-        9650 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 9650)
  AUTHORS   Bruning JC, Michael MD, Winnay JN, Hayashi T, Horsch D, Accili D, 
            Goodyear LJ, Kahn CR
  TITLE     A muscle-specific insulin receptor knockout exhibits features of the
            metabolic syndrome of NIDDM without altering glucose tolerance.
  JOURNAL   Mol Cell. 1998 Nov . 2(5):559-69.
  PUBMED    9844629
REFERENCE   2  (bases 1 to 9650)
  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. 
            1998 Nov . 2(5):559-69."
COMMENT     Sequence Label: pBS MCK promoter
FEATURES             Location/Qualifiers
     source          1..9650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     71..87
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6514..6530
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6564..6580)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6564..6580)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        6590..6608
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6658..6779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6695..6714)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6749..6768
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6862..6882)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6862..6880)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6901..6917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6901..6917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6914..6936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6925..6941
                     /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(6949..6955,6956..6973,6974..6979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6994..7015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7303..7891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7383..7402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8062..8853,8854..8922))
                     /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     8685..8704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8923..9027)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      9054..9510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9141..9160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9351..9372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(9636..9650,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(9650,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
       61 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcccc atcctggtct
      121 atagagagag ttccagaaca gccagggcta cagataaacc catctggaaa aacaaagttg
      181 aatgacccaa gaggggttct cagagggtgg cgtgtgctcc ctggcaagcc tatgacatgg
      241 ccggggcctg cctctctctg cctctgaccc tcagtggctc ccatgaactc cttgcccaat
      301 ggcatctttt tcctgcgctc cttgggttat tccagtctcc cctcagcatt ccttcctcag
      361 ggcctcgctc ttctctctgc tccctccttg cacagctggc tctgtccacc tcagatgtca
      421 cagtgctctc tcagaggagg aaggcaccat gtaccctctg tttcccaggt aagggttcaa
      481 tttttaaaaa tggttttttg tttgtttgtt tgtttgtttg tttgtttgtt tttcaagaca
      541 gggctcctct gtgtagtcct aactgtcttg aaactccctc tgtagaccag gtcgacctcg
      601 aactcttgaa acctgccacg gaccacccag tcaggtatgg aggtccctgg aatgagcgtc
      661 ctcgaagcta ggtgggtaag ggttcggcgg tgacaaacag aaacaaacac agaggcagtt
      721 tgaatctgag tgtattttgc agctctcaag caggggattt tatacataaa aaaaaaaaaa
      781 aaaaaaaaac caaacattac atctcttaga aactatatcc aatgaaacaa tcacagatac
      841 caaccaaaac cattgggcag agtaaagcac aaaaatcatc caagcattac aactctgaaa
      901 ccatgtattc agtgaatcac aaacagaaca ggtaacatca ttattaatat aaatcaccaa
      961 aatataacaa ttctaaaagg atgtatccag tgggggctgt cgtccaaggc tagtggcaga
     1021 tttccaggag caggttagta aatcttaacc actgaactaa ctctccagcc ccatggtcaa
     1081 ttattattta gcatctagtg cctaattttt ttttataaat cttcactatg taatttaaaa
     1141 ctattttaat tcttcctaat taaggctttc tttaccatat accaaaattc acctccaatg
     1201 acacacgcgt agccatatga aattttattg ttgggaaaat ttgtacctat cataatagtt
     1261 ttgtaaatga tttaaaaagc aaagtgttag ccgggcgtgg tggcacacgc ctttaatccc
     1321 tgcactcggg aggcaggggc aggaggattt ctgagtttga ggccagcctg gtctacagag
     1381 tgagttccag gacagccagg gctacacaga gaaaccctgt ctcgaacccc ccacccccca
     1441 aaaaaagcaa agtgttggtt tccttgggga taaagtcatg ttagtggccc atctctaggc
     1501 ccatctcacc cattattctc gcttaagatc ttggcctagg ctaccaggaa catgtaaata
     1561 agaaaaggaa taagagaaaa caaaacagag agattgccat gagaactacg gctcaatatt
     1621 ttttctctcc ggcgaagagt tccacaacca tctccaggag gcctccacgt tttgaggtca
     1681 atggcctcag tctgtggaac ttgtcacaca gatcttactg gaggtggtgt ggcagaaacc
     1741 cattcctttt agtgtcttgg gctaaaagta aaaggcccag aggaggcctt tgctcatctg
     1801 accatgctga caaggaacac gggtgccagg acagaggctg gaccccagga acaccttaaa
     1861 cacttcttcc cttctccgcc ccctagagca ggctcccctc accagcctgg gcagaaatgg
     1921 gggaagatgg agtgaagcca tactggctac tccagaatca acagagggag ccgggggcaa
     1981 tactggagaa gctggtctcc ccccaggggc aatcctggca cctcccaggc agaagaggaa
     2041 acttccacag tgcatctcac ttccatgaat cccctcctcg gactctgagg tccttggtca
     2101 cagctgaggt gcaaaaggct cctgtcatat tgtgtcctgc tctggtctgc cttccacagc
     2161 ttgggggcca cctagcccac ctctccctag ggatgagagc agccactacg ggtctaggct
     2221 gcccatgtaa ggaggcaagg cctggggaca cccgagatgc ctggttataa ttaacccaga
     2281 catgtggctg cccccccccc cccaacacct gctgcctgag cctcaccccc accccggtgc
     2341 ctgggtctta ggctctgtac accatggagg agaagctcgc tctaaaaata accctgtccc
     2401 tggtggatcc agggtgaggg gcaggctgag ggcggccact tccctcagcc gcaggtttgt
     2461 tttcccaaga atggtttttc tgcttctgta gcttttcctg tcaattctgc catggtggag
     2521 cagcctgcac tgggcttctg ggagaaacca aaccgggttc taacctttca gctacagtta
     2581 ttgcctttcc tgtagatggg cgactacagc cccaccccca cccccgtctc ctgtatcctt
     2641 cctgggcctg gggatcctag gctttcactg gaaatttccc cccaggtgct gtaggctaga
     2701 gtcacggctc ccaagaacag tgcttgcctg gcatgcatgg ttctgaacct ccaactgcaa
     2761 aaaatgacac ataccttgac ccttggaagg ctgaggcagg gggattgcca tgagtgcaaa
     2821 gccagactgg gtggcatagt tagaccctgt ctcaaaaaac caaaaacaat taaataacta
     2881 aagtcaggca agtaatccta ctcgggagac tgaggcagag ggattgttac atgtctgagg
     2941 ccagcctgga ctacataggg tttcaggcta gccctgtcta cagagtaagg ccctatttca
     3001 aaaacacaaa caaaatggtt ctcccagctg ctaatgctca ccaggcaatg aagcctggtg
     3061 agcattagca atgaaggcaa tgaaggaggg tgctggctac aatcaaggct gtgggggact
     3121 gagggcaggc tgtaacaggc ttgggggcca gggcttatac gtgcctggga ctcccaaagt
     3181 attactgttc catgttcccg gcgaagggcc agctgtcccc cgccagctag actcagcact
     3241 tagtttagga accagtgagc aagtcagccc ttggggcagc ccatacaagg ccatggggct
     3301 gggcaagctg cacgcctggg tccggggtgg gcacggtgcc cgggcaacga gctgaaagct
     3361 catctgctct caggggcccc tccctgggga cagcccctcc tggctagtca caccctgtag
     3421 gctcctctat ataacccagg ggcacagggg ctgcccccgg gtcaccacca cctccacagc
     3481 acagacagac actcaggagc cagccagcca ggtagggact gagagaaatc actggggtgg
     3541 gagtggggcg tgggagtcca agggtctgct cacccagtca tgttatggtt gtggattttg
     3601 cagcacaagt tgtggggaca aatgtctggg acacctaggt ctcaatagcc accaagtgtc
     3661 ccctccttgc aaggcagggt gggctggaac ttagtttagc agagttaatg gcccacacaa
     3721 agacagttgt ctcagtgaca cctgtcagtg gccctttaac tttgtaacca tgtggacctg
     3781 tgttgcagct ctgtgacctt gtgtctcact gtcctggtct gtctctatgt ctctctgtct
     3841 ctctgtctct atctctctct ttctgtctct ctctctccct ctctctttcg agatgggtca
     3901 gggggggtgg tgttctctgc atagccctgg ctgtcctgga actcactctg tagaccagcc
     3961 tggcctcgaa ctcagaaatc cacctgcctc ccaagtgctg ggattaaagg cgtgtgccac
     4021 caccgcccgg cgggtctttc ttgtgtgaga cttgggggct ctcactctta caggcccctg
     4081 gctttccttt gagtccttct gtctggctgt ctctgggatc ttgaaggcag gaaggactac
     4141 atgactcagt ttacctggag atcttagaga atctgtgatg agtttgggga ttccgaagct
     4201 ttctgcttct gcgtcttgcc tcggtgtcct gtctcctggg gtgcccctga gggagggggt
     4261 agcagaggat acagaacctt ctgaagggag agatctgggc tgggagcccg gggtgtcctt
     4321 gaggcccaga gcctggctgt gtgtcctcct ggccacccca gcccacctgt cccaatgctg
     4381 acttagtgca aggcgagcca gcaaggaggg aggacaggtg gcagtggggg gtgaggagca
     4441 tctaaaaata gccacaaagt agcagcttca agggctttgg gtctctgtct gccccacact
     4501 cttctctcag cttggtccac cttccctctc accttcctct gaggccccct tccagccccg
     4561 atggaggcct gatgtccccc atggtcagtg cttcagggat ctagtcaata aaattaataa
     4621 tgaaaaacaa cagtaataaa atacacgtga cgtgactggg gcagcttagg gcttagttca
     4681 aatcccagtg ttcacaccct ttaaaagaca agacaaaaca aaacagctgg ctgtggggga
     4741 gaacatcaga atccccctgg ggaggtgggg acaggggatc tgtggggctc catggccagg
     4801 cagcctagct ccaggcctgc gagagaccct acctcaagat aaaaataaaa taaaataaaa
     4861 taaaataaaa taaatatata aaataacaat gttgcagcac ctgaggtcac cactggaatg
     4921 tgcacacctg tgcacataca tgagcctgca ctacaaacaa aaatattaac agtaactgtt
     4981 agaatcccag ctgcaacttc atgccaggtg ccaggtccat gctcatcagt cagggactgg
     5041 aactcagaga tctcctggga aagcttcagt ctcacagatt caaaagccag agagatctag
     5101 tcacagcctg gggcccagag cagtgactta ggagagccgt gccttttaaa gtggaccttg
     5161 tagacagcca gaggtggagg gactgggaga agtggctgaa gcctccagac tcattcccac
     5221 gcccacatct ggactaattt ggatcagaat ctcaggggag cccttatggc ttttctcagg
     5281 tgtgcacata taatctttac cagggtcctc acacagagcc tgtcagattg gttttcaatt
     5341 tctgtgacaa acaccatgac caagacaatc tagaaaagag aaagcattaa tttggggctc
     5401 agggttctgg agcggcaggg aggtgggcat ggtgctggag cagaggctgg aagctcacat
     5461 ctttatcaac aaccagaggc agtgagagcc acttgggaat ggggtggctt ttcggaaatc
     5521 tcaaagccca caagcaatgg cacacctcct ccaacaaggc cacacctccg aatccttccc
     5581 aaacagttcc accgactggg gaccaaacat tcaaatatgt gagtctgagg ctcttctcat
     5641 tcaaatcacc acagacccaa gaacaatcga ataaaatatt tgtgttatgt gccaggcact
     5701 ggccgaggcg cttttcttgt cttttaatcc ctcccaagag gtcagcgatg ccacagtctc
     5761 catgttacag atgagtgaac aggaaagtca aacaggctcc tcagagtcac gcggctgctt
     5821 gtaagttgca aagccgaaat tcgaacccag accatctgat ccagatcctt tgctgctttt
     5881 attcatcttt ttattttatt ttattttatt ttaattcctg gtggcagggt ttctgtagcc
     5941 caggctaccc ttgaattcac tgcaatcctc ctgcctcagt ttcagagtgt tggaattaca
     6001 agcatggacc atcatgccca gttcctttgg gttgagatag agacctgtgt aggagcccag
     6061 actcgggctg gtctccagct ctctacgtag atgaagatga ccttgaactg ctgggatttc
     6121 aggcatgagc agccacaccc agatttgctg agcgccaaac tgttacccag ggtcctaagc
     6181 ttgctgggca agcactctgc cagcagaacc ccagccccag atcctgtatt tttgtagttg
     6241 tttttgttta tgtgactgtc cttttctggc tttagacaaa aggttttgcc ctccttttcc
     6301 agctagagag actgagtccc cagcaggatc acataggcag gatgtggcca catcaggcaa
     6361 cttgggctcc tgatgtttcc ttgcaaggct gaggttcaca gggggagaac cccccttttt
     6421 caagcccacg gtccgacgga ctgcaagccc ccagcaactg agttcttaag tctgaacccg
     6481 gtctgctcac agggtcccaa aggccgccgg cctcgaggtc gacggtatcg ataagcttga
     6541 tatcgaattc ctgcagcccg ggggatccac tagttctaga gcggccgcgt aatacgactc
     6601 actatagggc gaattcggat ccaagcttgg aattcgaatc agatcttatt aaagcagaac
     6661 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6721 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6781 catgtctggt cgactctaga ctcttccgct tcctcgctca ctgactcgag ctccaccgcg
     6841 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     6901 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6961 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7021 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7081 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7141 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7201 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7261 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7321 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7381 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7441 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7501 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7561 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7621 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7681 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7741 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7801 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7861 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7921 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7981 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8041 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8101 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8161 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8221 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8281 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8341 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8401 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8461 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8521 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8581 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8641 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8701 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8761 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8821 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8881 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8941 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9001 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     9061 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     9121 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     9181 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     9241 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     9301 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     9361 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9421 tttgccgatt tcggcctatt ggttaaaaaa atgagctgat ttaacaaaaa tttaacgcga
     9481 attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg
     9541 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9601 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
