LOCUS       62056_9455        4308 bp DNA     circular SYN 29-JUL-2019
DEFINITION  Cloning vector pENTR-BsaI-Tet LI, complete sequence.
ACCESSION   MK495753
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4308)
  AUTHORS   Chiasson D, Gimenez-Oya V, Bircheneder M, Bachmaier S, Studtrucker 
            T, Ryan J, Sollweck K, Leonhardt H, Boshart M, Dietrich P, Parniske 
            M.
  TITLE     A unified multi-kingdom Golden Gate cloning platform
  JOURNAL   Sci Rep 9 (1), 10131 (2019)
  PUBMED    31300661
REFERENCE   2  (bases 1 to 4308)
  AUTHORS   Chiasson D.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2019) Biology, LMU Munich, Grosshaderner Str. 2-4,
            Martinsried 82152, Germany
REFERENCE   3  (bases 1 to 4308)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep"; 
            date: "2019"; volume: "9"; issue: "1"; pages: "10131"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-FEB-2019) Biology, LMU Munich, Grosshaderner Str. 2-4, 
            Martinsried 82152, Germany"
FEATURES             Location/Qualifiers
     source          1..4308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             81..1268
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     rep_origin      1305..1893
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2223..2250)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(2342..2428)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     2492..2508
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2524..2623
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2687..2717
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2771..3427
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3772..4074
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(4142..4241)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(4259..4277)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgcattgaca gcttatcatc gataaactgt aatgcggtag tttatcacag ttaaattgct
       61 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg
      121 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg
      181 atatcgtcca ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt
      241 tgatgcaatt tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc
      301 cagtcctgct cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac
      361 ccgtcctgtg gattctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
      421 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
      481 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
      541 tgggcgccat ctccctgcac gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
      601 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgccgt ccgatgccct
      661 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgacc attgtggccg
      721 cacttatgac tgtgttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
      781 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
      841 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
      901 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
      961 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     1021 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     1081 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga
     1141 tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag
     1201 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca
     1261 cctcgacctg agacaattgt atccgaaaag atcaaaggat cttcttgaga tccttttttt
     1321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1441 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1681 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1921 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1981 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2041 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     2101 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     2161 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     2221 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     2281 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     2341 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     2401 cttgcgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
     2461 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
     2521 ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag
     2581 caatgctttt ttataatgcc aactttgtac aaaaaagcag gctccgcggc cgcccccttc
     2641 acctgagacc gcggcccctc gaggggtcgc attaggcacc ccaggcttta cactttatgc
     2701 ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa
     2761 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     2821 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     2881 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     2941 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat
     3001 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     3061 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     3121 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     3181 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     3241 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     3301 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     3361 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     3421 cggggcgtaa agatctggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
     3481 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
     3541 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
     3601 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
     3661 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
     3721 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
     3781 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     3841 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     3901 gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     3961 atgaccaccg atatggccag tgtgccggtt tccgttatcg gggaagaagt ggctgatctc
     4021 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     4081 tcaggctccc ttatacacag ccagtctgca ggtcgaccgg tctcaaaggg tgggcgcgcc
     4141 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     4201 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     4261 tatagtgagt cgtattacat ggtcatagcc agagctgcca ggaaacag
//
