LOCUS       Exported                7219 bp DNA     circular SYN 06-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7219)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Jun 15, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7219
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      1..195
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    539..679
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(865..1453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1699..2493)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MAREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
                     HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPA
                     KRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDL
                     FEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQ
                     PVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        2710..2912
                     /label=PvirB
     RBS             2922..2933
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2940..3659
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      3674..3745
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3746..3773
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3761..3779)
                     /label=pEA101-seq-R1
     CDS             5023..5652
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6081..7154
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
       61 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
      121 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
      181 cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac
      241 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
      301 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc
      361 gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg
      421 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      481 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
      541 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
      601 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
      661 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     1321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1501 aaactcacgt taagggattt tggtcatgca tgatatatct cccaatttgt gtagggctta
     1561 ttatgcacgc ttaaaaataa taaaagcaga cttgacctga tagtttggct gtgagcaatt
     1621 atgtgcttag tgcatctaac gcttgagtta agccgcgccg cgaagcggcg tcggcttgaa
     1681 cgaatttcta gctagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
     1741 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
     1801 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
     1861 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
     1921 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
     1981 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
     2041 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
     2101 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
     2161 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
     2221 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
     2281 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
     2341 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
     2401 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
     2461 tgatacttcg gcgataaccg cttcacgagc catggatccc tccctacaac tagtgatatc
     2521 gcccggagtt cgctcccaga aatatagcca tccctgcccc ctcacgtcaa tcccacgagc
     2581 ctcttatcca ttctcattga acgacggcga attgtattga ttggtgagct aatcaatatc
     2641 aaatatgaaa gggaaatcgt taaattattc aattattatc taattgaata atttaagaat
     2701 aatggtacct ccggttgctt agtcatggat gcgcggttct cggtccatgt tgcgttccaa
     2761 gacgccgggc gaggtttctc gcttcaattg aaatcataaa gaagcaattg aaaattttcg
     2821 agtaaccgac cctcccgata atcttcaaca taaaacaacg cacttcttcc aacgggagag
     2881 gcggtgttag ttgcgagcta aggagataag gtgaattcat taaagaggag aaaacgcgta
     2941 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3001 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3061 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3121 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3181 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3241 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3301 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3361 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3421 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3481 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3541 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3601 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     3661 gctttaagga tctcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3721 ctgttgtttg tcggtgaacg ctctccgcag aaaggcccac ccgaaggtga gccgcatgcc
     3781 aaccacaggg ttcccctcgg gatcaaagta ctttaaagta ctttaaagta ctttaaagta
     3841 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
     3901 cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
     3961 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
     4021 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
     4081 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
     4141 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga
     4201 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
     4261 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
     4321 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
     4381 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
     4441 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
     4501 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
     4561 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
     4621 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
     4681 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
     4741 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
     4801 cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt
     4861 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
     4921 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
     4981 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
     5041 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
     5101 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
     5161 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
     5221 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
     5281 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
     5341 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
     5401 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
     5461 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
     5521 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
     5581 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
     5641 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
     5701 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
     5761 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
     5821 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
     5881 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
     5941 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
     6001 ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc
     6061 cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg
     6121 cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac
     6181 gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca
     6241 aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg
     6301 acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca
     6361 gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga
     6421 tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca
     6481 gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc
     6541 gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg
     6601 tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct
     6661 gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc
     6721 gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg
     6781 aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca
     6841 cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg
     6901 gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat
     6961 ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt
     7021 tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg
     7081 cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat
     7141 ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag
     7201 gtcgaaaagg tctctttcc
//
