LOCUS       62056_21535        5686 bp DNA     circular SYN 28-AUG-2019
DEFINITION  Cloning vector pTrichoGate-6, complete sequence.
ACCESSION   MN007110
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5686)
  AUTHORS   Nogueira-Lopez G.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAY-2019) BPRC, Lincoln University, Ellesmere Jct Rd, 
            Lincoln, Christchurch, Canterbury 7647, New Zealand
REFERENCE   2  (bases 1 to 5686)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (31-MAY-2019) BPRC, Lincoln University, Ellesmere Jct Rd, Lincoln, 
            Christchurch, Canterbury 7647, New Zealand"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5686
                     /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"
     CDS             complement(1777..2493)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(2574..2592)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2599..2615)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2753..3052
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             3404..4195
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4405..4776
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      4917..5505
                     /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 gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttaagg tctctaagct tatacaactc
      361 gtccataccc aaggtgatac cagcagcggt gacaaactca aggaggacca tgtgatcacg
      421 cttctcgttg gggtccttag aaagagctga ctgagtggac agatagtggt tatctggcag
      481 gagaacaggg ccgtcaccga tgggagtgtt ttgttggtag tggtcggcga gttggacaga
      541 gccgtcttcg atgttatgtc ggattttgaa gttgacctta atgccgttct tctgcttgtc
      601 ggccataata taaacgttgt gggagttgta gttatactcc aacttatgac cgagaatgtt
      661 gccgtcctcc ttaaagtcaa tacccttcag ctcaatcctg ttgacgaggg tgtcaccttc
      721 aaacttgact tcagctcggg ttttgtaatt accatcatcc ttaaagaaga tggtacgttc
      781 ttgaacatag ccctctggca tagcgctctt aaagaaatcg tgctgcttca tgtgatcagg
      841 atagcggctg aagcactgta cgccgtaagt cagagttgta acgagggtcg gccaagggac
      901 gggcagcttg ccggtagtgc aaataaactt tagggtgagc ttgccgtagg ttgcatcgcc
      961 ttcaccctcg cctgagacgg agaacttgtg accgttgacg tcgccatcca actcgacaag
     1021 aatgggaacg acgccggtaa aaagttcctc gcccttgcta accttgtaca actcgtccat
     1081 gccaagggta ataccagcgg cggtgacgaa ctccaggaga accatgtggt cgcgcttctc
     1141 attggggtcc ttagacaggg cagactgagt tgagagatag tggttatcgg gcaggaggac
     1201 tggaccgtcg ccgatagggg tgttctgttg atagtggtcg gcgagttgaa cgctgccgtc
     1261 ctcgatgtta tgacggatct taaagttgac cttaatgccg ttcttctgtt tgtcagccat
     1321 aatataaacg ttatgagagt tatagttgta ttccaactta tgaccgagga tgttgccgtc
     1381 ctccttgaag tcaataccct taagctcgat tctgttgacg agggtgtcac cctcaaactt
     1441 aacttcagct cgggtcttgt agttaccgtc atccttaaag aagatagtac gctcttgaac
     1501 atagccctcg ggcatggcag acttaaagaa atcatgctgc ttcatgtgat ctggatagcg
     1561 tgagaagcac tggacgccgt aagtcagagt agtaacgagg gtaggccaag ggacgggcag
     1621 cttgccagtg gtgcaaatga acttcagggt gagcttgccg taggtggcgt cgccctcgcc
     1681 ctcgccgctg acagaaaact tgtggccgtt gacgtcgccg tccagttcga cgagaatagg
     1741 aacgacgccg gtgaagagtt cctcgccctt agaaaccttg tacagctcgt ccatgccgag
     1801 agtgatacca gcggcggtaa cgaactccag gaggaccatg tgatctcgct tctcgttggg
     1861 gtccttggac aaggcgctct gagtagaaag atagtggtta tcaggcagga ggacagggcc
     1921 gtcaccgata ggggtgtttt gctggtaatg atcagccagt tggacggagc cgtcctcaat
     1981 gttgtggcgg atcttgaagt tgaccttaat gccgttcttc tgcttatcgg ccatgatgta
     2041 gacgttatgt gagttatagt tgtactccaa cttgtggcca agaatgttac catcttcctt
     2101 aaagtcaatg cccttcagct cgatacggtt gacgagggtg tcaccttcga acttgacctc
     2161 agctcgggtc ttatagttgc catcgtcctt aaagaagatg gtgcgttcct ggacgtaacc
     2221 ctcaggcatg gcggacttaa agaaatcatg ttgcttcatg tggtcggggt agcgagaaaa
     2281 gcactgaacg ccgtaagtca gagtggtaac gagggttggc caagggacgg ggagcttgcc
     2341 agtggtgcag atgaacttca aggtgagctt gccgtaggta gcgtcgcctt caccctcgcc
     2401 gctaacggag aacttatgac cgttgacatc gccgtccaat tcgaccagga tgggaacgac
     2461 accagtgaac agctcttcgc cctttgatac cattagagac cttaagggcg aattctgcag
     2521 atatccatca cactggcggc cgctcgagca tgcatctaga gggcccaatt cgccctatag
     2581 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2641 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2701 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctatacgtac ggcagtttaa
     2761 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     2821 tattgacacg ccggggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     2881 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     2941 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     3001 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     3061 aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttcacgtaga aagccagtcc
     3121 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     3181 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     3241 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     3301 tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct gatggcgcag
     3361 gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3421 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3481 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3541 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3601 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3661 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3721 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3781 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3841 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3901 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3961 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4021 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4081 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4141 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat
     4201 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4261 acaccgcata caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4321 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4381 taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc ggtgctcacc
     4441 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     4501 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     4561 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     4621 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     4681 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     4741 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctaaa acttcatttt
     4801 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4861 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4921 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4981 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5041 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5101 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5161 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5221 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5281 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5341 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5401 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5461 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5521 gcctttttac ggttcctggg cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5581 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5641 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
