LOCUS       sequence-198180-        4544 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 4544)
  AUTHORS   Marigo V, Roberts DJ, Lee SM, Tsukurov O, Levi T, Gastier JM, 
            Epstein DJ, Gilbert DJ, Copeland NG, Seidman CE
  TITLE     Cloning, expression, and chromosomal location of SHH and IHH: two 
            human homologues of the Drosophila segment polarity gene hedgehog.
  JOURNAL   Genomics. 1995 Jul 1. 28(1):44-51.
  PUBMED    7590746
REFERENCE   2  (bases 1 to 4544)
  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: "Genomics. 
            1995 Jul 1. 28(1):44-51."
COMMENT     Sequence Label: pBS hShh (CT#401)
FEATURES             Location/Qualifiers
     source          1..4544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     67..83
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     68..84
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1704..1720)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1704..1720)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1757..1775)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1796..1812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1796..1812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1809..1831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1820..1836
                     /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(1844..1850,1851..1868,1869..1874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1889..1910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2027..2044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2198..2786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2278..2297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2957..3748,3749..3817))
                     /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     3580..3599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3818..3922)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3948..4403)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4085..4106)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4297..4316
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(4530..4544,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4544,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggtaccgggc
       61 cccccctcga ggtcgacggt atcgataagc ttgatatcga attccgggcg aggcagccag
      121 cgagggagag agcgagcggg cgagccggag cgaggaaggg aaagcgcaag agagagcgca
      181 cacgcacaca cccgccgcgc gcactcgcgc acggacccgc acggggacag ctcggaagtc
      241 atcagttcca tgggcgagat gctgctgctg gcgagatgtc tgctgctagt cctcgtctcc
      301 tcgctgctgg tatgctcggg actggcgtgc ggaccgggca gggggttcgg gaagaggagg
      361 caccccaaaa agctgacccc tttagcctac aagcagttta tccccaatgt ggccgagaag
      421 accctaggcg ccagcggaag gtatgaaggg aagatctcca gaaactccga gcgatttaag
      481 gaactcaccc ccaattacaa ccccgacatc atatttaagg atgaagaaaa caccggagcg
      541 gacaggctga tgactcagag gtgtaaggac aagttgaacg ctttggccat ctcggtgatg
      601 aaccagtggc caggagtgaa actgcgggtg accgagggct gggacgaaga tggccaccac
      661 tcagaggagt ctctgcacta cgagggccgc gcagtggaca tcaccacgtc tgaccgcgac
      721 cgcagcaagt acggcatgct ggcccgcctg gcggtggagg ccggcttcga ctgggtgtac
      781 tacgagtcca aggcacatat ccactgctcg gtgaaagcag agaactcggt ggcggccaaa
      841 tcgggaggct gcttcccggg ctcggccacg gtgcacctgg agcagggcgg caccaagctg
      901 gtgaaggacc tgagccccgg ggaccgcgtg ctggcggcgg acgaccaggg ccggctgctc
      961 tacagcgact tcctcacttt cctggaccgc gacgacggcg ccaagaaggt cttctacgtg
     1021 atcgagacgc gggagccgcg cgagcgcctg ctgctcaccg ccgcgcacct gctctttgtg
     1081 gcgccgcaca acgactcggc caccggggag cccgaggcgt cctcgggctc ggggccgcct
     1141 tccgggggcg cactggggcc tcgggcgctg ttcgccagcc gcgtgcgccc gggccagcgc
     1201 gtgtacgtgg tggccgagcg tgacggggac cgccggctcc tgcccgccgc tgtgcacagc
     1261 gtgaccctaa gcgaggaggc cgcgggcgcc tacgcgccgc tcacggccca gggcaccatt
     1321 ctcatcaacc gggtgctggc ctcgtgctac gcggtcatcg aggagcacag ctgggcgcac
     1381 cgggccttcg cgcccttccg cctggcgcac gcgctcctgg ctgcactggc gcccgcgcgc
     1441 acggaccgcg gcggggacag cggcggcggg gaccgcgggg gcggcggcgg cagagtagcc
     1501 ctaaccgctc caggtgctgc cgacgctccg ggtgcggggg ccaccgcggg catccactgg
     1561 tactcgcagc tgctctacca aataggcacc tggctcctgg acagcgaggc cctgcacccg
     1621 ctgggcatgg cggtcaagtc cagctgaagc cggggggccg ggggaggggc gcgggagggg
     1681 gccggaattc ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga
     1741 gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat
     1801 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1861 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1981 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2041 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2101 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2281 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2701 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2761 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2821 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2881 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2941 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3001 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3061 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3121 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3181 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3241 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3301 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3361 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3421 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3481 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3541 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3601 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3661 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3721 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3781 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3841 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3901 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     3961 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4021 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4081 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4141 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4201 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4261 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4321 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4381 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
     4441 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4501 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
