LOCUS       40924_34091        5887 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPasscode1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5887)
  AUTHORS   Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.
  TITLE     'Deadman' and 'Passcode' microbial kill switches for bacterial 
            containment
  JOURNAL   Nat. Chem. Biol. 12 (2), 82-86 (2016)
  PUBMED    26641934
REFERENCE   2  (bases 1 to 5887)
  AUTHORS   Chan CTY., Lee JW, Cameron DE, Bashor CJ, Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-OCT-2015) Department of Biological Engineering, 
            Massachusetts Institute of Technology, 77 Massachusetts Ave., 
            E25-302b, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 5887)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5887)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2016"; volume: "12"; issue: "2"; pages: "82-86"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-OCT-2015) Department of Biological Engineering, Massachusetts 
            Institute of Technology, 77 Massachusetts Ave., E25-302b, Cambridge,
            MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..115
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     CDS             122..1159
                     /codon_start=1
                     /product="GalR-LacI"
                     /label=GalR-LacI
                     /protein_id="ALP32097.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMESL
                     SYHPNANARALAQQTTETVGLVVGDVSDPFFGAMVKAVEQVAYHTGNFLLIGNGYHNEQ
                     KERQAIEQLIRHRCAALVVHAKMIPDADLASLMKQMPGMVLINRILPGFENRCIALDDR
                     YGAWLATRHLIQQGHTRIGYLCSNHSISDAEDRLQGYYDALAESGIAANDRLVTFGEPD
                     ESGGEQAMTELLGRGRNFTAVACYNDSMAAGAMGVLNDNGIDVPGEISLIGFDDVLVSR
                     YVRPRLTTVRYPIVTMATQAAELALALADNRPLPEITNVFSPTLVRRHSVSTPSLEASH
                     HATSD"
     terminator      1180..1274
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1323..1396
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     regulatory      1323..1370
                     /label=pLtetO-1
                     /note="pLtetO-1"
                     /regulatory_class="promoter"
     protein_bind    1323..1341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1348..1366
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      1377..1402
                     /label=RBS
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1403..2416
                     /codon_start=1
                     /product="CelR-LacI"
                     /label=CelR-LacI
                     /protein_id="ALP32098.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAIKEL
                     GYVPNRAARTLVTRRTDTVALVVSENNQKLFAEPFYAGIVLGVGVALSERGFQFVLATG
                     RSGIEHERLGGYLAGQHVDGVLLLSLHRDDPLPQMLDEAGVPYVYGGRPLGVPEEQVSY
                     VDIDNIGGGRQATQRLIETGHRRIATIAGPQDMVAGVERLQGYREALLAAGMEYDETLV
                     SYGDFTYDSGVAAMRELLDRAPDVDAVFAASDLMGLAALRVLRASGRRVPEDVAVVGYD
                     DSTVAEHAEPPMTSVNQPTELMGREMARLLVDRITGETTEPVRLVLETHLMVRESG"
     regulatory      2437..2531
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     terminator      2437..2531
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      complement(2571..2665)
                     /label=T0
                     /note="T0"
                     /regulatory_class="terminator"
     terminator      2571..2665
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2686..3783)
                     /codon_start=1
                     /product="LacI-ScrR"
                     /label=LacI-ScrR
                     /protein_id="ALP32099.1"
                     /translation="MKTKRVTIKDIAELAGVSKATASLVLNGRGKELRVAQETRERVLA
                     IMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMV
                     ERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINS
                     IIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAERE
                     GDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQ
                     TASPRALADSLMQLARQVSRLESGQ"
     regulatory      complement(3784..3809)
                     /label=RBS
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(3842..3870)
                     /label=pLlacO
                     /note="pLlacO"
                     /regulatory_class="promoter"
     terminator      3906..3992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(4156..4744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4832..4926)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4960..5751)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 gacgtctgtg caagtactag tatccatgga acgggcggcc gtccctatca gtgatagaga
       61 ttgacatccc tatcagtgat agagatactg agcacatcag caggacgcac tgaccggatc
      121 catgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
      181 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
      241 ggcgatggag tctcttagct atcacccgaa cgccaacgcc cgtgcgctgg cgcagcagac
      301 cactgaaacg gtcggtctgg tcgttggtga tgtttccgat ccgtttttcg gtgcaatggt
      361 gaaagcggtc gaacaggtgg cttatcacac cggtaatttt ttattgattg gcaacggtta
      421 ccacaacgaa caaaaagagc gtcaggccat tgagcaactg atccgccatc gctgtgctgc
      481 gttggtcgtc catgccaaaa tgatcccgga tgctgattta gcctcattaa tgaaacaaat
      541 gcccggtatg gtgctgatca accgtatcct gcctggcttt gaaaaccgtt gtattgctct
      601 ggacgatcgt tacggtgcct ggctggcaac gcgtcattta attcagcaag gtcatacccg
      661 cattggttat ctgtgctcta accactctat ttctgacgcc gaagatcgtc tgcaagggta
      721 ttacgatgcc cttgctgaaa gtggtattgc ggccaatgac cggctggtga catttggcga
      781 accagacgaa agcggcggcg aacaggcaat gaccgagctt ttgggacgag gaagaaattt
      841 cactgcggta gcctgttata acgattcaat ggcggcgggt gcgatgggcg ttctcaatga
      901 taatggtatt gatgtaccgg gtgagatttc gttaattggc tttgatgatg tgctggtgtc
      961 acgctatgtg cgtccgcgcc tgaccaccgt gcgttaccca atcgtgacga tggcgaccca
     1021 ggctgccgaa ctggctttgg cgctggcgga taatcgccct ctcccggaaa tcactaatgt
     1081 ctttagtccg acgctggtac gtcgtcattc agtgtcaact ccgtcgctgg aggcaagtca
     1141 tcatgcaacc agcgactaat gtacacgaga attggcttgg actcctgttg atagatccag
     1201 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     1261 ttattggtga gaatccaagc agtagtcacc gcgatctgaa gtatccatgg aacgggcggc
     1321 cgtccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacatca
     1381 gcaggacgca ctgaccggat ccatgaaacc agtaacgtta tacgatgtcg cagagtatgc
     1441 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     1501 aacgcgggaa aaagtggaag cggcgatcaa agagctgggc tacgtgccga accgcgcagc
     1561 ccgcaccctg gtcacccgac gtaccgacac cgtagccctg gtggtgtcgg aaaacaacca
     1621 gaagctcttc gccgaaccct tctatgccgg gatcgtgctc ggcgtggggg ttgctctgtc
     1681 cgaacgggga ttccagttcg tcctggccac gggccgctcc gggatagagc atgagcggct
     1741 gggcggctac ctggccggac agcacgtcga cggggtcctc ctgctgtcgc tccaccgcga
     1801 cgacccgctg ccgcagatgc tggacgaggc cggggtgccg tacgtctacg gcggccgtcc
     1861 gctcggcgtc cccgaagaac aggtgtccta tgtcgatatc gacaacatcg gcgggggacg
     1921 ccaggccacc cagcggctga tcgagaccgg gcaccggcgg atcgctacga tcgcgggccc
     1981 gcaggacatg gtcgctggtg tggaacgcct ccaggggtat cgcgaagcac tgctcgccgc
     2041 ggggatggag tacgacgaga cgctggtgag ctacggtgac ttcacctacg acagcggggt
     2101 ggccgcgatg cgggagctgc tggatcgggc ccccgacgtg gacgccgtgt tcgcggcctc
     2161 cgacttgatg gggctggccg cgctgcgggt gctgcgtgct tcgggacgcc gcgtgcccga
     2221 ggatgtggcg gtggtcggct acgacgactc gaccgtagcc gagcacgccg aaccgccgat
     2281 gaccagcgtc aaccagccca ccgagctgat gggccgggag atggcccggc tgctcgtgga
     2341 ccggatcacc ggggagacca ccgaaccggt gcggctggtg ctggagaccc atttgatggt
     2401 gcgggaatcc gggtgatgta cacgagaatt ggcttggact cctgttgata gatccagtaa
     2461 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     2521 ttggtgagaa tccaagcagt agtcaccgcg ctgcagtgac tactgcttgg attctcacca
     2581 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     2641 attactggat ctatcaacag gagtccaagc caattctcgc ccgggtcact gcccgctttc
     2701 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2761 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     2821 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     2881 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     2941 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     3001 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     3061 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     3121 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     3181 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     3241 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     3301 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     3361 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     3421 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     3481 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     3541 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     3601 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatgatcg ccagcacgcg
     3661 ctcgcgcgtc tcctgcgcca cgcgcagctc tttgccacgg ccgttgagca ccagactggc
     3721 ggtcgctttg gagacgcccg ccagttcggc gatatctttg atagttacgc gtttggtttt
     3781 catcttaaga tcctataaaa tggctagagc ctaggggtca gtgcgtcctg ctgatgtgct
     3841 cagtatcttg tgagcgctca caattgtcaa ctcgagagaa tcaatattaa tccaacgcgt
     3901 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     3961 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacttag gcgttcggct
     4021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4381 gtaggtcgtt cgctccaagc tgggctgtct gcacgaaccc cccgttcagc ccgaccgctg
     4441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4561 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4801 ttaagggatt ttggtcatgg ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     4861 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     4921 ggagtccaag ccaattctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4981 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     5041 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     5101 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     5161 catgatattc ggcaagcagg catcgccgtg ggtcacgacg agatcctcgc cgtcgggcat
     5221 acgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     5281 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     5341 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5401 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5461 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagccgc
     5521 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     5581 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgaccctgcg ctgacagccg
     5641 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5701 ctccacccaa gccgccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5761 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     5821 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg gcccaactgg
     5881 caattcc
//
