LOCUS       Exported                4700 bp ds-DNA     circular SYN 25-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4700)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 4, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Vector molecule:  253 with fake Ls
               Fragment ends: L2fake and L1fake
               Fragment size: 3185
            Insert molecule:  BC012986*
               Fragment ends: L1fake and L2fake
               Fragment size: 1515
FEATURES             Location/Qualifiers
     source          1..4700
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             668..2182
                     /codon_start=1
                     /label=tev-Hs.SMAD6
                     /note="SMAD family member 6"
                     /translation="ENLYFQGTMFRSKRSGLVRRLWRSRVVPDREEGGSGGGGGGDEDG
                     SLGSRAEPAPRAREGGGCGRSEVRPVAPRRPRDAVGQRGAQGAGRRRRAGGPPRPMSEP
                     GAGAGSSLLDVAEPGGPGWLPESDCETVTCCLFSERDAAGAPRDASDPLAGAALEPAGG
                     GRSREARSRLLLLEQELKTVTYSLLKRLKERSLDTLLEAVESRGGVPGGCVLVPRADLR
                     LGGQPAPPQLLLGRLFRWPDLQHAVELKPLCGCHSFAAAADGPTVCCNPYHFSRLCGPE
                     SPPPPYSRLSPRDEYKPLDLSDSTLSYTETEATNSLITAPGEFSDASMSPDATKPSHWC
                     SVAYWEHRTRVGRLYAVYDQAVSIFYDLPQGSGFCLGQLNLEQRSESVRRTRSKIGFGI
                     LLSKEPDGVWAYNRGEHPIFVNSPTLDAPGGRALVVRKVPPGYSIKVFDFERSGLQHAP
                     EPDAADGPYDPNSVRISFAKGWGPCYSRQFITSCPCWLEILLNNPR"
     CDS             668..688
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2181..2280)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2298..2316)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2321..2337)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2768..3559
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            3681..3790
                     /note="incP origin of transfer"
     rep_origin      4050..4638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcgaa aacctgtact tccaaggcac catgttcagg tccaaacgct cggggctggt
      721 gcggcgactt tggcgaagtc gtgtggtccc cgaccgggag gaaggcggca gcggcggcgg
      781 cggtggcggc gacgaggatg ggagcttggg cagccgagct gagccggccc cgcgggcaag
      841 agagggcgga ggctgcggcc gctccgaagt ccgcccggta gccccgcggc ggccccggga
      901 cgcagtggga cagcgaggcg cccagggcgc ggggaggcgc cggcgcgcag ggggcccccc
      961 gaggcccatg tcggagccag gggccggcgc tgggagctcc ctgctggacg tggcggagcc
     1021 gggaggcccg ggctggctgc ccgagagtga ctgcgagacg gtgacctgct gtctcttttc
     1081 ggagcgggac gccgccggcg cgccccggga cgccagcgac cccctggccg gggcggccct
     1141 ggagccggcg ggcggcgggc ggagtcgcga agcgcgctcg cggctgctgc tgctggagca
     1201 ggaactcaaa accgtcacgt actcgctgct gaagcggctc aaggagcgct cgctggacac
     1261 gctgctggag gcggtggagt cccgcggcgg cgtgccgggc ggctgcgtgc tggtgccgcg
     1321 cgccgacctc cgcctgggcg gccagcccgc gccgccgcag ctgctgctcg gccgcctctt
     1381 tcgctggccc gacctgcagc acgccgtgga gctgaagccc ctgtgcggct gccacagctt
     1441 cgccgccgcc gccgacggcc ctaccgtgtg ctgcaacccc taccacttca gccggctctg
     1501 cgggcccgaa tctccgccac ctccctactc tcggctgtct cctcgcgacg agtacaagcc
     1561 actggatctg tccgattcca cattgtctta cactgaaacg gaggctacca actccctcat
     1621 cactgctccg ggtgaattct cagacgccag catgtctccg gacgccacca agccgagcca
     1681 ctggtgcagc gtggcgtact gggagcaccg gacgcgcgtg ggccgcctct atgcggtgta
     1741 cgaccaggcc gtcagcatct tctacgacct acctcagggc agcggcttct gcctgggcca
     1801 gctcaacctg gagcagcgca gcgagtcggt gcggcgaacg cgcagcaaga tcggcttcgg
     1861 catcctgctc agcaaggagc ccgacggcgt gtgggcctac aaccgcggcg agcaccccat
     1921 cttcgtcaac tccccgacgc tggacgcgcc cggcggccgc gccctggtcg tgcgcaaggt
     1981 gccccccggc tactccatca aggtgttcga cttcgagcgc tcgggcctgc agcacgcgcc
     2041 cgagcccgac gccgccgacg gcccctacga ccccaacagc gtccgcatca gcttcgccaa
     2101 gggctggggg ccctgctact cccggcagtt catcacctcc tgcccctgct ggctggagat
     2161 cctcctcaac aaccccagat aatcaacttt cttgtacaaa gttggcatta taagaaagca
     2221 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2281 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2341 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2401 atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc
     2461 cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc
     2521 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     2581 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     2641 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     2701 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     2761 aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     2821 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     2881 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
     2941 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     3001 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     3061 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     3121 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     3181 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     3241 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     3301 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     3361 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     3421 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     3481 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     3541 caaggtagtc ggcaaataac cctcgactct agaggatccc cgggtaccga gctcgaattc
     3601 cgtgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc
     3661 atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc
     3721 caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc
     3781 tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca
     3841 aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc
     3901 gaagcagggt tatgcagcgg aaaacggaat tcgagctcgg tacccgggga tcctctagag
     3961 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     4021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4441 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4681 tggccttttg ctcacatgtt
//
