LOCUS       62056_17825        6122 bp DNA     circular SYN 04-SEP-2019
DEFINITION  Plant binary expression vector pNC-Green0029-35S, complete sequence.
ACCESSION   MK896904
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Yan P, Zeng Y, Shen W, Tuo D, Li X, Zhou P.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-MAY-2019) Institute of Tropical Bioscience and 
            Biotechnology, Chinese Academy of Tropical Agriculture Sciences, 
            Xueyuan Road, Haikou, Hainan 571101, China
REFERENCE   2  (bases 1 to 6122)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (07-MAY-2019) Institute of Tropical Bioscience and Biotechnology, 
            Chinese Academy of Tropical Agriculture Sciences, Xueyuan Road, 
            Haikou, Hainan 571101, China"
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        184..529
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     promoter        725..827
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             858..1160
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1251..1503
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1521..1537)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1574..1592)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1613..1629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1637..1653)
                     /label=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)."
     promoter        complement(1661..1692)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1707..1728)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    1967..1991
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2082..2670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2774..3586)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3877..4312
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4445..4467
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     terminator      complement(4491..4742)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4785..5576)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5610..5789)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     6036..6052
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6059..6077
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6103..6119
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catggagtca aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca
       61 gttcatacag agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga
      121 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
      181 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
      241 tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca
      301 ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg
      361 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
      421 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
      481 gcaagaccct tcctctatat aaggaagttc atttcatttg gagagaacac gggggactct
      541 tgacagtggt ctctgtccag tcctggcctc gtcggccatt ctcgactaag ttggcagcat
      601 cacccgacgc actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa
      661 taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag
      721 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
      781 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaactt ttgctgacga
      841 gaacagggac tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc
      901 gtctgtttgt ggatgtacag agtgatatta ttgacacgcc tgggcgacgg atggtgatcc
      961 ccctggccag tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc
     1021 atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtatccg
     1081 ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta
     1141 acctgatgtt ctggggaata taaatgtcag gctcccttat acacagggcc agtctggcca
     1201 cttgtggtct gctgagaccg tgtgaattgg tgaccagctc gaatttcccc gatcgttcaa
     1261 acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca
     1321 tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat
     1381 ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa
     1441 acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag
     1501 atcgggcctg cagcccgggg gatccactag ttctagagcg gccgccaccg cggtggagct
     1561 ccagcttttg ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc
     1621 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaaghc
     1681 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1921 ttatccacag aatcagggga taacgcagga aagaacatga aggccttgac aggatatatt
     1981 ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag
     2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2521 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggttac aaccaattaa
     2761 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     2821 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     2881 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     2941 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     3001 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     3061 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     3121 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     3181 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     3241 aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta
     3301 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     3361 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     3421 gtttcagaaa caactctggc gcatcgggct tcccatacaa tccatagatt gtcgcacctg
     3481 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3541 ttaatcgcgg cctggagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     3601 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     3661 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
     3721 agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa
     3781 agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca
     3841 ctttctggct ggatgatggg gcgattcagg cgatccccat ccaacagccc gccgtcgagc
     3901 gggctttttt atccccggaa gcctgtggat agagggtagt tatccacgtg aaaccgctaa
     3961 tgccccgcaa agccttgatt cacggggctt tccggcccgc tccaaaaact atccacgtga
     4021 aatcgctaat cagggtacgt gaaatcgcta atcggagtac gtgaaatcgc taataaggtc
     4081 acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa tagccctttc agatcaacag
     4141 cttgcaaaca cccctcgctc cggcaagtag ttacagcaag tagtatgttc aattagcttt
     4201 tcaattatga atatatatat caattattgg tcgcccttgg cttgtggaca atgcgctacg
     4261 cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc accgtcgagc gccagcgcct
     4321 ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta taaatttttt tttttgaaaa
     4381 agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga tttccgatcc ccggaattag
     4441 atcttggcag gatatattgt ggtgtaacgt tatcagcttg catgccggtc gatctagtaa
     4501 catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat
     4561 cgcgtattaa atgtataatt gcgggactct aatcaaaaaa cccatctcat aaataacgtc
     4621 atgcattaca tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc
     4681 atcgcaagac cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacga
     4741 tctgcttgac tctagctaga gtccgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     4801 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     4861 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     4921 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     4981 tccaccatga tattcggcaa gcaggcatcg ccctgggtca cgacgagatc ctcgccgtcg
     5041 ggcatccgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     5101 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtcctcg ctcgatgcga
     5161 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     5221 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     5281 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     5341 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttggagt
     5401 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     5461 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     5521 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcct
     5581 cgatcgagtt gagagtgaat atgagactct aattggatac cgaggggaat ttatggaacg
     5641 tcagtggagc atttttgaca agaaatattt gctagctgat agtgacctta ggcgactttt
     5701 gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca gaaacccgcg
     5761 gctgagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc
     5821 gcgtcatcgg cgggggtcat aacgtgactc ccttaattct catgtatcga taacattaac
     5881 gtttacaatt tcgcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5941 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6001 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
     6061 atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg acggtatcga
     6121 ta
//
