LOCUS       sequence-324115-        4242 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 4242)
  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 4242)
  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 hIhh (CT#402)
FEATURES             Location/Qualifiers
     source          1..4242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        223..241
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     278..294
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     278..294
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1612..1628)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1613..1629)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1653..1672)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1654..1672)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1679..1696)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1679..1695)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1688..1710)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1837..2292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1924..1943)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2134..2155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2318..2422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2423..2491,2492..3283)
                     /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(2641..2660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3454..4042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3943..3962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4196..4213
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag
      241 ggaacaaaag ctggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg
      301 gctgcaggaa ttccggcgcc tcatgaccca gcgctgcaag gaccgcctga actcgctggc
      361 tatctcggtg atgaaccagt ggcccggtgt gaagctgcgg gtgaccgagg gctgggacga
      421 ggacggccac cactcagagg agtccctgca ttatgagggc cgcgcggtgg acatcaccac
      481 atcagaccgc gaccgcaata agtatggact gctggcgcgc ttggcagtgg aggccggctt
      541 tgactgggtg tattacgagt caaaggccca cgtgcattgc tccgtcaagt ccgagcactc
      601 ggccgcagcc aagacgggcg gctgcttccc tgccggagcc caggtacgcc tggagagtgg
      661 ggcgcgtgtg gccttgtcag ccgtgaggcc gggagaccgt gtgctggcca tgggggagga
      721 tgggagcccc accttcagcg atgtgctcat tttcctggac cgcgagcccc acaggctgag
      781 agccttccag gtcatcgaga ctcaggaccc cccacgccgc ctggcactca cacccgctca
      841 cctgctcttt acggctgaca atcacacgga gccggcagcc cgcttccggg ccacatttgc
      901 cagccacgtg cagcctggcc agtacgtgct ggtggctggg gtgccaggcc tgcagcctgc
      961 ccgcgtggca gctgtctcta cacacgtggc cctcggggcc tacgccccgc tcacaaagca
     1021 tgggacactg gtggtggagg atgtggtggc atcctgcttc gcggccgtgg ctgaccacca
     1081 cctggctcag ttggccttct ggcccctgag actctttcac agcttggcat ggggcagctg
     1141 gaccccgggg gagggtgtgc attggtaccc ccagctgctc taccgcctgg ggcgtctcct
     1201 gctagaagag ggcagcttcc acccactggg catgtccggg gcagggagct gaaaggactc
     1261 caccgctgcc ctcctggaac tgctgtactg ggtccagaag cctctcagcc aggagggagc
     1321 tggccctgga agggacctga gctgggggac actggctcct gccatctcct ctgccatgaa
     1381 gatacaccat tgagacttga ctgggcaaca ccagcgtccc ccacccgcgt cgtggtgtag
     1441 tcatagagct gcaagctgag ctggcgaggg gatggttgtt gacccctctc tcctagagac
     1501 cttgaggctg gcacggcgac tcccaactca gcctgctctc actacgagtt ttcatactct
     1561 gcctccccca ttggggaggg cccattcccc ggaattcgat atcaagctta tcgataccgt
     1621 cgacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat tacaattcac
     1681 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1741 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1801 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     1861 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1921 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1981 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2041 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2101 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2161 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2221 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2281 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2341 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2401 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2461 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2521 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2581 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2641 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2701 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2761 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2821 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2881 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2941 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3001 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3061 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3121 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3181 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3241 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3301 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3361 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3421 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3481 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3541 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3601 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3661 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3721 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3781 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3841 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3901 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3961 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4021 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4081 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4141 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4201 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa cc
//
