LOCUS       Exported                4607 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 4607)
  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:  BC009682*
               Fragment ends: L1fake and L2fake
               Fragment size: 1422
FEATURES             Location/Qualifiers
     source          1..4607
                     /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..2089
                     /codon_start=1
                     /label=tev-Hs.SMAD5
                     /note="SMAD family member 5"
                     /translation="ENLYFQGTMTSMASLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDA
                     LVKKLKKKKGAMEELEKALSSPGQPSKCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWP
                     DLQSHHELKPLDICEFPFGSKQKEVCINPYHYKRVESPVLPPVLVPRHNEFNPQHSLLV
                     QFRNLSHNEPHMPQNATFPDSFHQPNNTPFPLSPNSPYPPSPASSTYPNSPASSGPGSP
                     FQLPADTPPPAYMPPDDQMGQDNSQPMDTSNNMIPQIMPSISSRDVQPVAYEEPKHWCS
                     IVYYELNNRVGEAFHASSTSVLVDGFTDPSNNKSRFCLGLLSNVNRNSTIENTRRHIGK
                     GVHLYYVGGEVYAECLSDSSIFVQSRNCNFHHGFHPTTVCKIPSSCSLKIFNNQEFAQL
                     LAQSVNHGFEAVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKV
                     LTQMGSPLNPISSVS"
     CDS             668..688
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2088..2187)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2205..2223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2228..2244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2675..3466
                     /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            3588..3697
                     /note="incP origin of transfer"
     rep_origin      3957..4545
                     /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 catgacgtca atggccagct tgttttcttt
      721 tactagtcca gcagtaaagc gattgttggg ctggaaacaa ggtgatgagg aggagaaatg
      781 ggcagaaaag gcagttgatg ctttggtgaa gaaactaaaa aagaaaaagg gtgccatgga
      841 ggaactggag aaagccttga gcagtccagg acagccgagt aaatgtgtca ctattcccag
      901 atctttagat ggacgcctgc aggtttctca cagaaaaggc ttaccccatg ttatatattg
      961 tcgtgtttgg cgctggccgg atttgcagag tcatcatgag ctaaagccgt tggatatttg
     1021 tgaatttcct tttggatcta agcaaaaaga agtttgtatc aacccatacc actataagag
     1081 agtggagagt ccagtcttac ctccagtatt agtgcctcgt cataatgaat tcaatccaca
     1141 acacagcctt ctggttcagt ttaggaacct gagccacaat gaaccacaca tgccacaaaa
     1201 tgccacgttt ccagattctt tccaccagcc caacaacact ccttttccct tatctccaaa
     1261 cagcccttat cccccttctc ctgctagcag cacatatccc aactccccag caagttctgg
     1321 accaggaagt ccatttcagc tcccagctga tacgcctcct cctgcctata tgccacctga
     1381 tgatcagatg ggtcaagata attcccagcc tatggataca agcaataata tgattcctca
     1441 gattatgccc agtatatcca gcagggatgt tcagcctgtt gcctatgaag agcctaaaca
     1501 ttggtgttca atagtctact atgaattaaa caatcgtgtt ggagaagctt ttcatgcatc
     1561 ttctactagt gtgttagtag atggattcac agatccttca aataacaaaa gtagattctg
     1621 cttgggtttg ttgtcaaatg ttaatcgtaa ttcgacaatt gaaaacacta ggcgacatat
     1681 tggaaaaggt gttcatctgt actatgttgg tggagaggtg tatgcggaat gcctcagtga
     1741 cagcagcata tttgtacaga gtaggaactg caactttcat catggctttc atcccaccac
     1801 tgtctgtaag attcccagca gctgcagcct caaaattttt aacaatcagg agtttgctca
     1861 gcttctggct caatctgtca accatgggtt tgaggcagta tatgagctca ccaaaatgtg
     1921 taccattcgg atgagttttg tcaagggttg gggagcagaa tatcaccggc aggatgtaac
     1981 cagcacccca tgttggattg agattcatct tcatgggcct cttcagtggc tggataaagt
     2041 ccttactcag atgggctccc ctctgaaccc catatcttct gtttcataat caactttctt
     2101 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2161 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2221 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2281 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     2341 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     2401 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     2461 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     2521 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     2581 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     2641 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     2701 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     2761 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     2821 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     2881 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     2941 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     3001 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     3061 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     3121 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     3181 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     3241 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     3301 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     3361 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     3421 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgactctaga
     3481 ggatccccgg gtaccgagct cgaattccgt gataggtggg ctgcccttcc tggttggctt
     3541 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
     3601 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
     3661 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
     3721 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
     3781 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa acggaattcg
     3841 agctcggtac ccggggatcc tctagagtcg agccacccat gaccaaaatc ccttaacgtg
     3901 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4081 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4321 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     4381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
