LOCUS       62056_21450        6990 bp DNA     circular SYN 28-AUG-2019
DEFINITION  Cloning vector pTrichoGate-1, complete sequence.
ACCESSION   MN007105
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  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 6990)
  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..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    67..88
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        103..133
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..157
                     /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     165..181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(194..250)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(251..267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    647..671
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1971..2597
                     /codon_start=1
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3034..4098
                     /codon_start=1
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      4167..4361
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    4705..4845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5031..5619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5709..6500)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6925..6949
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 acatagagac cgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
       61 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      121 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      181 ccatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga
      241 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      301 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      361 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
      421 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
      481 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      541 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacg gtctcattgt
      601 aatgggtcat ctatgttact agatcgggaa ttaaactatc agtgtttgac aggatatatt
      661 ggcgggtaaa cctaagagaa aagagcgttt attagaataa cggatattta aaagggcgtg
      721 aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga
      781 tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct gacgttcagt
      841 gcagccgtct tctgaaaacg acatgtcgca caagtcctaa gttacgcgac aggctgccgc
      901 cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt agaatacttg
      961 cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc tgctgggcta
     1021 tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac tgcacgcggc
     1081 cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc gcccggagct
     1141 ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca ggctagaccg
     1201 cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg aggccggcgc
     1261 gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg gccgcatggt
     1321 gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg accgcacccg
     1381 gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc ctaccctcac
     1441 cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca ccgtgaaaga
     1501 ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg agcgcagcga
     1561 ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg cattgaccga
     1621 ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat gaaaccgcac
     1681 caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga gatgatcgcg
     1741 gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg tgcggctgca tgaaatcctg
     1801 gccggtttgt ctgatgccaa gctggcggcc tggccggcca gcttggccgc tgaagaaacc
     1861 gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc
     1921 gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc atgaaggtta
     1981 tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc catctagccc
     2041 gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc cagggcagtg
     2101 cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc
     2161 cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg atcgacggag
     2221 cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc gtgctgattc
     2281 cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag ctggttaagc
     2341 agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca
     2401 aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag ctgcccattc
     2461 ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg
     2521 ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa
     2581 ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa gcacaaacac
     2641 gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc
     2701 agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc acaccaagct
     2761 gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg aatacatcgc
     2821 gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta gcggctaaag
     2881 gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg
     2941 gaggaacggg cggttggcca ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca
     3001 ctggaacccc caagcccgag gaatcggcgt gacggtcgca aaccatccgg cccggtacaa
     3061 atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag
     3121 cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga
     3181 atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc
     3241 aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat
     3301 agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc
     3361 gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc
     3421 atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc
     3481 atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt
     3541 gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta
     3601 gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag
     3661 aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta
     3721 aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc
     3781 gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc
     3841 gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa
     3901 gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag
     3961 ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag
     4021 gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc
     4081 gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg
     4141 gaaaaaggtc gaaaaggtac ctttcctgtg gatagcacgt acattgggaa cccaaagccg
     4201 tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca
     4261 cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta
     4321 aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca
     4381 gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct
     4441 tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat
     4501 ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc
     4561 accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4621 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4681 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4741 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4801 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     4861 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4921 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4981 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5041 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5101 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5161 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5221 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5281 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5341 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5401 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5461 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5521 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5581 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5641 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct
     5701 aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg
     5761 atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc
     5821 gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca
     5881 ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg
     5941 gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga
     6001 tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc
     6061 agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat
     6121 atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg
     6181 ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga
     6241 ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc
     6301 agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg
     6361 tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga
     6421 gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt
     6481 gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac
     6541 cccaagaagc taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc
     6601 ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat
     6661 cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca
     6721 atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt
     6781 tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct
     6841 gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
     6901 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
     6961 tagacaactt aataacacat tgcggattgg
//
