LOCUS       Exported                4448 bp ds-DNA     circular SYN 05-AUG-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4448)
  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:  BC014690*
               Fragment ends: L1fake and L2fake
               Fragment size: 1263
FEATURES             Location/Qualifiers
     source          1..4448
                     /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..1930
                     /codon_start=1
                     /label=tev-Mm.Tgfb3 C227S/C229S
                     /note="transforming growth factor, beta 3, C227S/C229S"
                     /translation="ENLYFQGTMKMHLQRALVVLALLNLATISLSLSTCTTLDFGHIKK
                     KRVEAIRGQILSKLRLTSPPEPSVMTHVPYQVLALYNSTRELLEEMHGEREEGCTQETS
                     ESEYYAKEIHKFDMIQGLAEHNELAVCPKGITSKVFRFNVSSVEKNGTNLFRAEFRVLR
                     VPNPSSKRTEQRIELFQILRPDEHIAKQRYIGGKNLPTRGTAEWLSFDVTDTVREWLLR
                     RESNLGLEISIHSPSHTFQPNGDILENVHEVMEIKFKGVDNEDDHGRGDLGRLKKQKDH
                     HNPHLILMMIPPHRLDSPGQGSQRKKRALDTNYCFRNLEENCCVRPLYIDFRQDLGWKW
                     VHEPKGYYANFCSGPCPYLRSADTTHSTVLGLYNTLNPEASASPCCVPQDLEPLTILYY
                     VGRTPKVEQLSNMVVKSCKCS"
     CDS             668..688
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1929..2028)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2046..2064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2069..2085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2516..3307
                     /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            3429..3538
                     /note="incP origin of transfer"
     rep_origin      3798..4386
                     /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 catgaagatg cacttgcaaa gggctctggt
      721 agtcctggcc ctgctgaact tggccacaat cagcctctct ctgtccactt gcaccacgtt
      781 ggacttcggc cacatcaaga agaagagggt ggaagccatt aggggacaga tcttgagcaa
      841 gctcaggctc accagccccc ctgagccatc ggtgatgacc cacgtcccct atcaggtcct
      901 ggcactttac aacagcaccc gggagttgct ggaagagatg cacggggaga gggaggaagg
      961 ctgcactcag gagacctcgg agtctgagta ctatgccaaa gagatccata aattcgacat
     1021 gatccagggg ctggcggagc acaatgaact ggccgtttgc cccaaaggaa ttacctctaa
     1081 ggtttttcgt ttcaatgtgt cctcagtgga gaaaaatgga accaatctgt tccgggcaga
     1141 gttccgggtc ttgcgggtgc ccaaccccag ctccaagcgc acagaacaga gaattgagct
     1201 cttccagata cttcgaccgg atgagcacat agccaagcag cgctacatag gtggcaagaa
     1261 tctgcccaca aggggcaccg ctgagtggct gtctttcgat gtcactgaca ctgtgcgcga
     1321 gtggctgttg aggagagagt ccaacttggg tctggaaatc agcatccaca gtccaagtca
     1381 cacctttcag cccaatggag acatactgga aaatgttcat gaggtgatgg aaatcaaatt
     1441 caaaggagtg gacaatgaag atgaccatgg ccgtggagac ctggggcgtc tcaagaagca
     1501 gaaggatcac cacaacccac acctgatcct catgatgatc cccccacacc gactggacag
     1561 cccaggccag ggtagtcaga ggaagaagag ggccctggac accaattact gcttccgcaa
     1621 cctggaggag aactgctgtg tacgccccct ttatattgac ttccggcagg atctaggctg
     1681 gaaatgggtc cacgaaccta agggttacta tgccaacttc tgctcaggcc cttgcccata
     1741 cctccgcagc gcagacacaa cccatagcac ggtgcttgga ctatacaaca ccctgaaccc
     1801 agaggcgtct gcctcgccat gctgcgtccc ccaggacctg gagcccctga ccatcttgta
     1861 ctatgtgggc agaaccccca aggtggagca gctgtccaac atggtggtga agtcgtgtaa
     1921 gtgcagctaa tcaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1981 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2041 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2101 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     2161 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     2221 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     2281 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2341 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     2401 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2461 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     2521 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     2581 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     2641 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     2701 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     2761 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     2821 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     2881 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     2941 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3001 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3061 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3121 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3181 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3241 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3301 caaataaccc tcgactctag aggatccccg ggtaccgagc tcgaattccg tgataggtgg
     3361 gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc
     3421 ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata
     3481 agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg
     3541 ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata
     3601 tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta
     3661 tgcagcggaa aacggaattc gagctcggta cccggggatc ctctagagtc gagccaccca
     3721 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     3781 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3901 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3961 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4021 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4081 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4141 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     4201 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4261 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4321 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4381 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4441 cacatgtt
//
