LOCUS       Exported                4693 bp ds-DNA     circular SYN 15-DEC-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTBC0086 pBb(B5)1k-GFPuv
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4693)
  AUTHORS   Taylor Cook
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 5, 2018 from SnapGene 3.1.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4693
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..6
                     /note="AatII"
     promoter        7..84
                     /gene="lacI (mutant)"
                     /note="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     -35_signal      21..26
                     /note="-35"
     -10_signal      44..49
                     /note="-10"
     CDS             85..1167
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     misc_feature    1399..1428
                     /note="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1436..1452
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     misc_feature    1458..1463
                     /note="EcoRI"
     misc_feature    1467..1472
                     /note="BglII"
     CDS             1493..2209
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /note="GFPuv"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    2210..2215
                     /note="BamHI"
     misc_feature    2219..2224
                     /note="XhoI"
     terminator      2242..2313
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2329..2356
                     /note="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     misc_feature    2363..2368
                     /note="AvrII"
     CDS             complement(2544..3051)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /note="B5 Rep"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQP"
     misc_feature    3052..3625
                     /note="oriV-mutant"
     misc_feature    3073
                     /note="G Insertion"
     misc_feature    3482
                     /note="C Insertion"
     terminator      3638..3732
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(3763..4557)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4646..4693)
                     /note="NEOKAN_promoter"
ORIGIN
        1 gacgtcgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
       61 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
      121 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
      181 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
      241 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
      301 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
      361 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
      421 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
      481 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
      541 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
      601 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
      661 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
      721 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
      781 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
      841 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
      901 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
      961 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     1021 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agcgcgaatt
     1201 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     1261 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     1321 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     1381 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
     1441 agcggataac aatttcagaa ttcaaaagat cttttaagaa ggagatatac atatgagtaa
     1501 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1561 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1621 ccttaaattt atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac
     1681 tttctcttat ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt
     1741 tttcaagagt gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga
     1801 cgggaactac aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat
     1861 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt ctcggacaca aactggagta
     1921 caactataac tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc
     1981 taacttcaaa attcgccaca acattgaaga tggctccgtt caactagcag accattatca
     2041 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2101 acaatctgcc ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt
     2161 tgtaactgct gctgggatta cacatggcat ggatgagctc tacaaataag gatccaaact
     2221 cgagtaagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2281 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     2341 ggtgggcctt tctgcgttta tacctaggct acagccgata gtctggaaca gcgcacttac
     2401 gggttgctgc gcaacccaag tgctaccggc gcggcagcgt gacccgtgtc ggcggctcca
     2461 acggctcgcc atcgtccaga aaacacggct catcgggcat cggcaggcgc tgctgcccgc
     2521 gccgttccca ttcctccgtt tcggtcaagg ctggctgggc ggctcctcgc cggggccggt
     2581 cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc catgccccaa
     2641 cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca ccgacaggcg
     2701 caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg ccaacacggt
     2761 gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt ccttgactgc
     2821 gtattggacc gtccgcaaag aacgtccgat gagcttggaa agtgtcttct ggctgaccac
     2881 cacggcgttc tggtggccca tctgcgccac gaggtgatgc agcagcattg ccgccgtggg
     2941 tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt gcacccagtg
     3001 accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca tgcttatctc
     3061 catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca acttttcggc
     3121 agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt ttctttaacc
     3181 tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc ccttttttaa
     3241 gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg tgcccaaaga
     3301 agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg gcaaaaagtg
     3361 gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag gtggcgctgc
     3421 ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg cgggcttcgc
     3481 ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa ggccaagccg
     3541 ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac cggcaagcga
     3601 agcgcgcagg ccgcaggccg gaggcactag tgcttggatt ctcaccaata aaaaacgccc
     3661 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     3721 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
     3781 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3841 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3901 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3961 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     4021 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     4081 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     4141 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     4201 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     4261 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4321 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     4381 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4441 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4501 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     4561 aaacgatcct catcctgtct cttgatcaga tcatgatccc ctgcgccatc agatccttgg
     4621 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     4681 agctggcaat tcc
//
