LOCUS       40924_48368        3356 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZero++ Kan, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3356)
  AUTHORS   Wu G, Zhu Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2004) Fish Genetics, Institute of Hydrobiology, 
            Chinese Academy of Science, 7# South Road of East Lake, Wuhan, Hubei
            430072, People's Republic of China
REFERENCE   2  (bases 1 to 3356)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3356)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (09-MAR-2004) Fish Genetics, Institute of Hydrobiology, Chinese 
            Academy of Science, 7# South Road of East Lake, Wuhan, Hubei 430072,
            People's Republic of China"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(449..467)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(474..490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             628..927
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     rep_origin      955..1334
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1385..2176)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2587..3175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcagatc tcatatgcct aggagctccg
      301 cggacgtcga caagcttggt accaagtatg acatggcgcc atgggagtct tggggatcca
      361 ctagtaacgg ccgccagtgt gctggaattc tgcagatatc catcacactg gcggccgctc
      421 gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc
      481 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
      541 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
      601 caacagttgc gcagcctata cgtacggcag tttaaggttt acacctataa aagagagagc
      661 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgccggg gcgacggatg
      721 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
      781 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
      841 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
      901 gccattaacc tgatgttctg gggaatataa atgtcaggcc tgaatggcga atggacgcgc
      961 cctgtagcgg cgcattaagc gcgcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1021 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1081 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagagcttt
     1141 acggcacctc gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     1201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1261 gttccaaact ggaacaacac tcaaccctat cgcggtctat tcttttgatt tataagggat
     1321 gttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat tttaacaaaa
     1381 ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     1441 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     1501 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     1561 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccgtgggtca
     1621 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
     1681 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     1741 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     1801 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     1861 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     1921 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     1981 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     2041 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     2101 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     2161 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat cctgatcccc
     2221 tgcgccatca gtccttggcg gcgagaaagc catccagttt actttgcagg gcttcccaac
     2281 cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca
     2341 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     2401 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     2461 actggctttc tacgtgaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2521 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2581 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2641 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2701 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2941 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3001 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3061 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3121 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3181 cagcaacgcg gcctttttac ggttcctggg cttttgctgg ccttttgctc acatgttctt
     3241 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3301 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
