LOCUS       sequence-343687-        3398 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 3398)
  AUTHORS   Hartmann C, Tabin CJ
  TITLE     Wnt-14 plays a pivotal role in inducing synovial joint formation in 
            the developing appendicular skeleton.
  JOURNAL   Cell. 2001 Feb 9. 104(3):341-51.
  PUBMED    11239392
REFERENCE   2  (bases 1 to 3398)
  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: "Cell. 2001 
            Feb 9. 104(3):341-51."
COMMENT     Sequence Label: pGEM cGdf5 (CT#688)
FEATURES             Location/Qualifiers
     source          1..3398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      496..505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      500..509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(539..558)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(540..558)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(565..582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(565..581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(574..596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      722..1177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(809..828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1019..1040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1255..1359
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1360..1428,1429..2220)
                     /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(1578..1597)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2391..2979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2880..2899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3133..3150
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3267..3288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3303..3308,3309..3326,3327..3333)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3341..3357
                     /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     3346..3368
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3365..3381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3365..3381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atttaggtga cactatagaa tactcaagct atgcatccaa cgcgttggga gctctcccat
       61 atggtcgacc tgcaggcggc cgcgaattca ctagtgattt gggaggtctt cgacatctgg
      121 aagcttttta ggaactttaa aaacttggtt aacttgtgtt tcgaactgga aacttttgac
      181 agggggagag cggttgatct caggactgtg ggatttaaca gaacgggaag acaggtcaac
      241 gaaaaggctc tcttcttggt atttgggagg acaaagaaaa gggacttatt tttcaatgaa
      301 atcaaagcta gatccggcca ggacgacaaa accgtgtatg ggtacttatt caatcagagg
      361 cggaagagaa gagctcctct ggcaacacgg caagggaaga ggcccagtaa gaacctgaag
      421 gcaaggtgta gcagaaaagc cctgcatgtg aattttaagg atatggggtg ggatgattgg
      481 aatcgaattc ccgcggccgc catggcggcc gggagcatgc gacgtcgggc ccaattcgcc
      541 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      601 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      661 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      721 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      841 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1081 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1141 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     1201 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     1261 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1321 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1381 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1441 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1501 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1561 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1621 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1681 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1741 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1801 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1861 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1921 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1981 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2041 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2101 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2161 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2221 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2281 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2521 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3181 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3241 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     3301 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     3361 cacacaggaa acagctatga ccatgattac gccaagct
//
