LOCUS       Exported                4759 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 4759)
  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:  hTGFBR1
               Fragment ends: blunt
               Fragment size: 1574
FEATURES             Location/Qualifiers
     source          1..4759
                     /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             687..2222
                     /codon_start=1
                     /label=tev-Hs.TGFBR1
                     /translation="ENLYFQGTMEAAVAAPRPRLLLLVLAAAAAAAAALLPGATALQCF
                     CHLCTKDNFTCVTDGLCFVSVTETTDKVIHNSMCIAEIDLIPRDRPFVCAPSSKTGSVT
                     TTYCCNQDHCNKIELPTTVKSSPGLGPVELAAVIAGPVCFVCISLMLMVYICHNRTVIH
                     HRVPNEEDPSLDRPFISEGTTLKDLIYDMTTSGSGSGLPLLVQRTIARTIVLQESIGKG
                     RFGEVWRGKWRGEEVAVKIFSSREERSWFREAEIYQTVMLRHENILGFIAADNKDNGTW
                     TQLWLVSDYHEHGSLFDYLNRYTVTVEGMIKLALSTASGLAHLHMEIVGTQGKPAIAHR
                     DLKSKNILVKKNGTCCIADLGLAVRHDSATDTIDIAPNHRVGTKRYMAPEVLDDSINMK
                     HFESFKRADIYAMGLVFWEIARRCSIGGIHEDYQLPYYDLVPSDPSVEEMRKVVCEQKL
                     RPNIPNRWQSCEALRVMAKIMRECWYANGAARLTALRIKKTLSQLSQQEGIKM"
     CDS             687..707
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2240..2339)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2357..2375)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2380..2396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2827..3618
                     /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            3740..3849
                     /note="incP origin of transfer"
     rep_origin      4109..4697
                     /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 agttggcttt gtacaaaaaa gttggcgaaa acctgtactt ccaaggcacc atggaggcgg
      721 cggtcgctgc tccgcgtccc cggctgctcc tcctcgtgct ggcggcggcg gcggcggcgg
      781 cggcggcgct gctcccgggg gcgacggcgt tacagtgttt ctgccacctc tgtacaaaag
      841 acaattttac ttgtgtgaca gatgggctct gctttgtctc tgtcacagag accacagaca
      901 aagttataca caacagcatg tgtatagctg aaattgactt aattcctcga gataggccgt
      961 ttgtatgtgc accctcttca aaaactgggt ctgtgactac aacatattgc tgcaatcagg
     1021 accattgcaa taaaatagaa cttccaacta ctgtaaagtc atcacctggc cttggtcctg
     1081 tggaactggc agctgtcatt gctggaccag tgtgcttcgt ctgcatctca ctcatgttga
     1141 tggtctatat ctgccacaac cgcactgtca ttcaccatcg agtgccaaat gaagaggacc
     1201 cttcattaga tcgccctttt atttcagagg gtactacgtt gaaagactta atttatgata
     1261 tgacaacgtc aggttctggc tcaggtttac cattgcttgt tcagagaaca attgcgagaa
     1321 ctattgtgtt acaagaaagc attggcaaag gtcgatttgg agaagtttgg agaggaaagt
     1381 ggcggggaga agaagttgct gttaagatat tctcctctag agaagaacgt tcgtggttcc
     1441 gtgaggcaga gatttatcaa actgtaatgt tacgtcatga aaacatcctg ggatttatag
     1501 cagcagacaa taaagacaat ggtacttgga ctcagctctg gttggtgtca gattatcatg
     1561 agcatggatc cctttttgat tacttaaaca gatacacagt tactgtggaa ggaatgataa
     1621 aacttgctct gtccacggcg agcggtcttg cccatcttca catggagatt gttggtaccc
     1681 aaggaaagcc agccattgct catagagatt tgaaatcaaa gaatatcttg gtaaagaaga
     1741 atggaacttg ctgtattgca gacttaggac tggcagtaag acatgattca gccacagata
     1801 ccattgatat tgctccaaac cacagagtgg gaacaaaaag gtacatggcc cctgaagttc
     1861 tcgatgattc cataaatatg aaacattttg aatccttcaa acgtgctgac atctatgcaa
     1921 tgggcttagt attctgggaa attgctcgac gatgttccat tggtggaatt catgaagatt
     1981 accaactgcc ttattatgat cttgtacctt ctgacccatc agttgaagaa atgagaaaag
     2041 ttgtttgtga acagaagtta aggccaaata tcccaaacag atggcagagc tgtgaagcct
     2101 tgagagtaat ggctaaaatt atgagagaat gttggtatgc caatggagca gctaggctta
     2161 cagcattgcg gattaagaaa acattatcgc aactcagtca acaggaaggc atcaaaatgt
     2221 aatcaacttt cttgtacaaa gtcaactttc ttgtacaaag ttggcattat aagaaagcat
     2281 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     2341 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     2401 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca
     2461 tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc
     2521 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
     2581 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
     2641 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     2701 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     2761 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     2821 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     2881 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     2941 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
     3001 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     3061 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     3121 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     3181 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     3241 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     3301 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     3361 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     3421 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     3481 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     3541 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     3601 aaggtagtcg gcaaataacc ctcgactcta gaggatcccc gggtaccgag ctcgaattcc
     3661 gtgataggtg ggctgccctt cctggttggc ttggtttcat cagccatccg cttgccctca
     3721 tctgttacgc cggcggtagc cggccagcct cgcagagcag gattcccgtt gagcaccgcc
     3781 aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct
     3841 atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa
     3901 aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg
     3961 aagcagggtt atgcagcgga aaacggaatt cgagctcggt acccggggat cctctagagt
     4021 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     4081 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4141 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4201 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4261 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4321 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4381 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4441 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4501 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4561 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4621 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4681 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4741 ggccttttgc tcacatgtt
//
