LOCUS       Exported                4592 bp ds-DNA    circular SYN 19-SEP-2018
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pLXMA (RK2) Level 2A
SOURCE      synthetic DNA construct
  ORGANISM  PCR product
REFERENCE   1  (bases 1 to 4592)
  AUTHORS   SynthSys Center
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 31, 2020 from SnapGene 2.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4592
                     /organism="PCR product"
                     /mol_type="other DNA"
     misc_feature    1..24
                     /note="assembly linker"
                     /note="color: #a6acb3"
     regulatory      25..72
                     /note="color: #ffffff"
     misc_feature    133..157
                     /note="LB"
                     /note="color: #a6acb3"
     misc_feature    180..210
                     /note="LB"
                     /note="color: #a6acb3"
     misc_feature    184..208
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
                     /note="color: #ffe4c4"
     misc_feature    223..228
                     /note="BioBrick prefix"
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
                     /note="color: #99ccff"
     misc_feature    236..239
                     /note="GGAG Overlap"
                     /note="color: #ff0000"
     promoter        251..285
                     /note="J23106 promoter"
                     /note="color: #00ffff"
     RBS             294..305
                     /note="B0034 RBS"
                     /note="color: #ffff00"
     CDS             312..1031
                     /codon_start=1
                     /note="sfGFP"
                     /note="color: #ffff00"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     YYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK*"
     terminator      1036..1107
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     terminator      1123..1150
                     /note="T7Te terminator"
                     /note="phage T7 early transcription terminator"
                     /note="color: #ffffff"
     misc_feature    1162..1165
                     /note="ACCC Overlap"
                     /note="color: #ff0000"
     misc_feature    1172..1181
                     /note="RB from pTi6"
                     /note="color: #a6acb3"
     misc_feature    1196..1219
                     /note="RB"
                     /note="color: #a6acb3"
     regulatory      complement(1284..1324)
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    1334..1357
                     /note="assembly linker"
                     /note="color: #a6acb3"
     gene            complement(1358..2263)
                     /gene="aadA"
                     /note="aadA"
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(1358..2263)
                     /codon_start=1
                     /gene="aadA"
                     /product="spectinomycin adenyltransferase"
                     /note="aadA"
                     /note="AadA"
                     /note="color: #993366"
                     /protein_id="ASG92153.1"
                     /translation="MPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIA
                     EVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRAL
                     INDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPA
                     TIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDER
                     NVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRA
                     DQLEEFVHYVKGEITKVVGK"
     CDS             complement(1358..2149)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /note="SmR"
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    2347..2370
                     /note="assembly linker"
                     /note="color: #a6acb3"
     gene            complement(2421..3569)
                     /gene="trfA"
                     /note="trfA"
                     /note="color: #a6acb3; direction: LEFT"
     CDS             complement(2421..3569)
                     /codon_start=1
                     /gene="trfA"
                     /product="replication protein TrfA"
                     /note="trfA"
                     /note="color: #993366"
                     /protein_id="ASG92154.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(2820..2822)
                     /codon_start=1
                     /note="A250V 20 fold copy number"
                     /note="color: #993366"
                     /translation="A"
     rep_origin      3962..4493
                     /note="oriV"
                     /note="incP origin of replication"
                     /note="color: #ffff00"
ORIGIN
        1 ccatcatcag ttcggtggtc ttccgacgaa caataaggcc gcaaatcgcg gcctttttta
       61 ttgataacaa aaccggctca gttctgcgta gaaaccaaca tgcaagctcc accgggtgca
      121 aagcggcagc ggcggcagga tatattcaat tgtaaatggc ttcatgtccg ggaaatctac
      181 tggtggcagg atatattgtg gtgtaaacaa tggagaaaaa gagaattcgg tctcaggaga
      241 tatgcaggtg tttacggcta gctcagtcct aggtatagtg ctagctacta gagaaagagg
      301 agaaatacta aatgcgtaaa ggcgaagaac tgttcacggg cgtagttccg attctggtcg
      361 agctggacgg cgatgtgaac ggtcataagt ttagcgttcg cggtgaaggt gagggcgacg
      421 cgaccaacgg caaactgacc ctgaagttca tctgcaccac cggtaaactg ccggtgcctt
      481 ggccgacctt ggtgacgacg ttgacgtatg gcgtgcagtg ttttgcgcgt tatccggacc
      541 acatgaaaca acacgatttc ttcaaatctg cgatgccgga gggttacgtc caggagcgta
      601 ccatttcctt caaggatgat ggctactaca aaactcgcgc agaggttaag tttgaaggtg
      661 acacgctggt caatcgtatc gaattgaagg gtatcgactt taaagaggat ggtaacattc
      721 tgggccataa actggagtat aacttcaaca gccataatgt ttacattacg gcagacaagc
      781 aaaagaacgg catcaaggcc aatttcaaga ttcgccacaa tgttgaggac ggtagcgtcc
      841 aactggccga ccattaccag cagaacaccc caattggtga cggtccggtt ttgctgccgg
      901 ataatcacta tctgagcacc caaagcgtgc tgagcaaaga tccgaacgaa aaacgtgatc
      961 acatggtcct gctggaattt gtgaccgctg cgggcatcac ccacggtatg gacgagctgt
     1021 ataagtaatg agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1081 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     1141 cctttctgcg cacctgcata taccctgcag gaattgttga ttttgtgatg actgatggca
     1201 ggatatatgc ggttgtaatt catttttatt gtctaaattt ctgtatttgt ttgtttgttc
     1261 ggttgtaaat ttttttggaa gaacaagaaa agaaaaaaca cccgttaggg tgtttttagt
     1321 tagtgtggcg cgccgacttg cgacatgcgg tcctttgtta tttgccgact accttggtga
     1381 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
     1441 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
     1501 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
     1561 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
     1621 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
     1681 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
     1741 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
     1801 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
     1861 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
     1921 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
     1981 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
     2041 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
     2101 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcatg atgtttaact
     2161 ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac atcaaacatc
     2221 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt accccaaaaa
     2281 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     2341 aggttcgatc ggattggcgg ttatgcggtt gcgatgctgg tggctgctga acccccagcc
     2401 ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg
     2461 tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac
     2521 ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg
     2581 tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg
     2641 cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc
     2701 tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg
     2761 tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc
     2821 gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag
     2881 cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc
     2941 ttcgcgtact ccaagacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg
     3001 ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc
     3061 tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc
     3121 atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg
     3181 atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cttcgctgac ctgttttgcc
     3241 aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc
     3301 atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc
     3361 gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta
     3421 gcttgctgga ctatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg
     3481 cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg
     3541 tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccggaatta
     3601 attccccgga tcgatccgtc gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     3661 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     3721 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     3781 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     3841 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg gctgccattt
     3901 ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt
     3961 tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg
     4021 cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa
     4081 aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg
     4141 cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc
     4201 tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac
     4261 cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag
     4321 gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc
     4381 ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct
     4441 catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccctacatg
     4501 gctctgctgt agtgagtggg ttgcgctccg gcagcggtcc tgatcccccg cagaaaaaaa
     4561 ggatctcaag aagatccttt gatcttttct ac
//
