LOCUS       40924_8896       10516 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10516)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 10516)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..720
                     /label=ECFP
                     /note="enhanced CFP"
     terminator      752..1004
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1026..1050
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2350..2976
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             3413..4477
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4546..4740
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    5084..5224
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5410..5998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6088..6879)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    7304..7328
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(7406..7580)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7623..8645)
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     promoter        complement(8713..9390)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    9581..9602
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9617..9647
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9655..9671
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9679..9695
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9705..9761
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(9765..9781)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10158..10503
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
       61 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      121 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      181 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg
      241 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      301 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      361 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      421 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag
      481 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      541 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      601 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
      661 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
      721 taaattacag gtgaccagct cgaatttccc cgatcgttca aacatttggc aataaagttt
      781 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
      841 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
      901 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
      961 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat taaactatca
     1021 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataac
     1081 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac
     1141 cacagggttc ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag
     1201 tcggcttctg acgttcagtg cagccgtctt ctgaaaacga catgtcgcac aagtcctaag
     1261 ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc
     1321 gcataaagta gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg
     1381 ccgctggcct gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac
     1441 gggccgaact gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca
     1501 ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga
     1561 cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc
     1621 gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca
     1681 cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc
     1741 taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg
     1801 gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg
     1861 aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc
     1921 gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc
     1981 gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg
     2041 aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat
     2101 cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt
     2161 gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag
     2221 cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa
     2281 cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa
     2341 ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc
     2401 atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat
     2461 tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc
     2521 gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac
     2581 ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca
     2641 gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac
     2701 ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt
     2761 gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc
     2821 gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact
     2881 gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc
     2941 caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa
     3001 tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg
     3061 caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg
     3121 cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc
     3181 tgctatctga atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga
     3241 tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc
     3301 gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgtc
     3361 tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg acggtcgcaa
     3421 accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa
     3481 ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg
     3541 gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc
     3601 gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta
     3661 tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa
     3721 gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt
     3781 ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt
     3841 ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg
     3901 cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa
     3961 gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca
     4021 gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat
     4081 tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct
     4141 agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca
     4201 ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg
     4261 cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag
     4321 cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct
     4381 gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta
     4441 ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg
     4501 gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctc tttcctgtgg atagcacgta
     4561 cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc
     4621 gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt
     4681 ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata
     4741 actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg
     4801 ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc
     4861 tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg
     4921 ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     4981 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5041 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     5101 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     5161 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5221 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5281 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5341 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5401 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5461 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5521 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5581 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5641 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5701 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5761 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5821 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5881 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5941 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6061 gattttggtc atgcattcta ggtactaaaa caattcatcc agtaaaatat aatattttat
     6121 tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat actgttcttc
     6181 cccgatatcc tccctgatcg accggacgca gaaggcaatg tcataccact tgtccgccct
     6241 gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa agatgttgct
     6301 gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg tctttaaaaa
     6361 atcatacagc tcgcgcggat ctttaaatgg agtgtcttct tcccagtttt cgcaatccac
     6421 atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt ctaagctatt
     6481 cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact ccgcatacag
     6541 ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa ggacgccatc
     6601 ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca ggacctttgg
     6661 aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca catcataggt
     6721 ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt ctcccaccag
     6781 cttatatacc ttagcaggag acattccttc cgtatctttt acgcagcggt atttttcgat
     6841 cagttttttc aattccggtg atattctcat tttagccatt tattatttcc ttcctctttt
     6901 ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc caattcactg
     6961 ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt gttttcaaag
     7021 ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc acaggcagca
     7081 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     7141 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     7201 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
     7261 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     7321 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
     7381 gaattaacgc cgaattaatt cgggggatct ggattttagt actggatttt ggttttagga
     7441 attagaaatt ttattgatag aagtatttta caaatacaaa tacatactaa gggtttctta
     7501 tatgctcaac acatgagcga aaccctatag gaaccctaat tcccttatct gggaactact
     7561 cacacattat tatggagaaa ctcgagcttg tcgatcgaca gatccggtcg gcatctactc
     7621 tatttctttg ccctcggacg agtgctgggg cgtcggtttc cactatcggc gagtacttct
     7681 acacagccat cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc
     7741 ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa
     7801 ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat atacgcccgg
     7861 agtcgtggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc
     7921 atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg ggaatccccg
     7981 aacatcgcct cgctccagtc aatgaccgct gttatgcggc cattgtccgt caggacattg
     8041 ttggagccga aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc
     8101 atcagctcat cgagagcctg cgcgacggac gcactgacgg tgtcgtccat cacagtttgc
     8161 cagtgataca catggggatc agcaatcgcg catatgaaat cacgccatgt agtgtattga
     8221 ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg
     8281 atcgcatcca tagcctccgc gaccggttgt agaacagcgg gcagttcggt ttcaggcagg
     8341 tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat aggtcaggct ctcgctaaac
     8401 tccccaatgt caagcacttc cggaatcggg agcgcggccg atgcaaagtg ccgataaaca
     8461 taacgatctt tgtagaaacc atcggcgcag ctatttaccc gcaggacata tccacgccct
     8521 cctacatcga agctgaaagc acgagattct tcgccctccg agagctgcat caggtcggag
     8581 acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt
     8641 ttcatatctc attgcccccc gggatctgcg aaagctcgag agagatagat ttgtagagag
     8701 agactggtga tttcagcgtg tcctctccaa atgaaatgaa cttccttata tagaggaagg
     8761 tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa
     8821 tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt
     8881 gggggtccat ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta
     8941 tcgcaatgat ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga
     9001 cagatagctg ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc
     9061 tcaatagccc tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg
     9121 tcgtgctcca ccatgttatc acatcaatcc acttgctttg aagacgtggt tggaacgtct
     9181 tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag
     9241 gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc
     9301 ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt
     9361 cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct
     9421 ttgatattct tggagtagac gagagtgtcg tgctccacca tgttggcaag ctgctctagc
     9481 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     9541 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     9601 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     9661 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgaattc gagctcggta
     9721 cccggggatc ctctagagtc gacctgcagg catgcaagct tggcactggc cgtcgtttta
     9781 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     9841 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     9901 cgcagcctga atggcgaatg ctagagcagc ttgagcttgg atcagattgt cgtttcccgc
     9961 cttcagttta gcttcatgga gtcaaagatt caaatagagg acctaacaga actcgccgta
    10021 aagactggcg aacagttcat acagagtctc ttacgactca atgacaagaa gaaaatcttc
    10081 gtcaacatgg tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca
    10141 gaagaccaaa gggcaattga gacttttcaa caaagggtaa tatccggaaa cctcctcgga
    10201 ttccattgcc cagctatctg tcactttatt gtgaagatag tggaaaagga aggtggctcc
    10261 tacaaatgcc atcattgcga taaaggaaag gccatcgttg aagatgcctc tgccgacagt
    10321 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
    10381 acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa
    10441 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagaga
    10501 acacggggga ctcttg
//
