LOCUS       40924_2832        6671 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector p4eng3VT-lacI, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Boehm CR, Grant PK, Haseloff J.
  TITLE     Programmed emergence of a domain of gene expression in a bacterial 
            population
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6671)
  AUTHORS   Boehm CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-OCT-2016) Plant Sciences, University of Cambridge, 
            Downing Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom
REFERENCE   3  (bases 1 to 6671)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6671)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-OCT-2016) Plant Sciences, University of Cambridge, Downing 
            Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..966
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     RBS             1244..1266
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1278..1994
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
     terminator      2017..2088
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2104..2131
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      2138..2172
                     /label=PJ23101*
                     /note="PJ23101*"
                     /regulatory_class="promoter"
     RBS             2181..2192
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2208..3287
                     /label=lacI
                     /note="lac repressor"
     CDS             3288..3320
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      complement(3371..3414)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3455..3482
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3489..3509
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             3613..4470
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      4484..4543
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     terminator      complement(4603..4632)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    4679..4700
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     terminator      complement(4707..4734)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4750..4821)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4844..5560)
                     /label=mTurquoise2
                     /note="enhanced monomeric variant of CFP (Goedhart et al., 
                     2012)"
     RBS             5567..5578
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(5587..5621)
                     /label=pJ23101
                     /note="pJ23101"
                     /regulatory_class="promoter"
     misc_feature    5622..5642
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5678..5709
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     terminator      5834..5928
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6420..6642
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ctagaccaat tgagatgtat gtctgaatta gttgttttca aagcaaatga actagcgatt
       61 agtcgctatg acttaacgga gcatgaaacc aagctaattt tatgctgtgt ggcactactc
      121 aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac ttataaccaa
      181 tacgctcaga tgatgaacat cagtagggaa aatgcttatg gtgtattagc taaagcaacc
      241 agagagctga tgacgagaac tgtggaaatc aggaatcctt tggttaaagg ctttgagatt
      301 ttccagtgga caaactatgc caagttctca agcgaaaaat tagaattagt ttttagtgaa
      361 gagatattgc cttatctttt ccagttaaaa aaattcataa aatataatct ggaacatgtt
      421 aagtcttttg aaaacaaata ctctatgagg atttatgagt ggttattaaa agaactaaca
      481 caaaagaaaa ctcacaaggc aaatatagag attagccttg atgaatttaa gttcatgtta
      541 atgcttgaaa ataactacca tgagtttaaa aggcttaacc aatgggtttt gaaaccaata
      601 agtaaagatt taaacactta cagcaatatg aaattggtgg ttgataagcg aggccgcccg
      661 actgatacgt tgattttcca agttgaacta gatagacaaa tggatctcgt aaccgaactt
      721 gagaacaacc agataaaaat gaatggtgac aaaataccaa caaccattac atcagattcc
      781 tacctacata acggactaag aaaaacacta cacgatgctt taactgcaaa aattcagctc
      841 accagttttg aggcaaaatt tttgagtgac atgcaaagta agtatgatct caatggttcg
      901 ttctcatggc tcacgcaaaa acaacgaacc acactagaga acatactggc taaatacgga
      961 aggatctgag gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa
     1021 aagtagaaca actgttcacc gttacatatc aaagggaaaa ctgtccatat gcacagatga
     1081 aaacggtgta aaaaagatag atacatcaga gcttttacga gtttttggtg cattcaaagc
     1141 tgttcaccat gaacagatcg acaatgtaac tactagagta ctagaggcga aattaatacg
     1201 actcactgta gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga
     1261 aggagatata ctactagatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1321 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     1381 agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc
     1441 ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct
     1501 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1561 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1621 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1681 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg
     1741 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg
     1801 gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1861 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     1921 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1981 acgagctgta caagtaataa tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg
     2041 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     2101 actggctcac cttcgggtgg gcctttctgc gtttatattt acagctagct cagtcctagg
     2161 ttttatgcta gctactagag aaagaggaga aatactagat ggtgaatgtg aaaccagtaa
     2221 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     2281 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     2341 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     2401 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     2461 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     2521 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     2581 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     2641 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     2701 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     2761 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     2821 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     2881 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     2941 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     3001 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     3061 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     3121 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     3181 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3241 cattaatgca gctggcacga caggtttccc gactggaaag cgggcaggct gcaaacgacg
     3301 aaaactacgc tttagtagct taataactct gatagtgcta gtgtagatct ctactagagc
     3361 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     3421 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     3481 cgtttatata ctagtagcgg ccgctgcagt gatctacact agcactatca gtgctaaata
     3541 cattcaaata tctatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3601 aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgccgaagat
     3721 cagttgggtg cacgtgtggg ttacatcgaa ctggacctca acagcggtaa gattcttgag
     3781 agttttcgcc ccgaagaacg tttcccaatg atgagcactt ttaaagttct gctctgtggc
     3841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga cggcatgaca
     3961 gtacgcgaat tatgcagcgc tgccataacc atgagtgata acacggcggc caacttactt
     4021 ctgacaacga tcggaggacc gaaggagctt accgcttttt tgcacaacat gggtgatcat
     4081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4141 gacaccacga tgcctgtagc tatggcaaca acgttgcgca aactcttaac tggcgaactt
     4201 cttactctcg cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4321 gagcgtgggt cccgcggtat tattgcagcc ctggggccag atggtaagcc ctcccgtatc
     4381 gtagttatct acacgacggg gagccaggca actatggacg aacgtaatcg ccagatcgct
     4441 gagataggtg cctccctgat taagcattgg taataatact agctccggca aaaaaacggg
     4501 caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgtttgcca
     4561 cctgacgtct aagaaaagga atattcagca atttgcccgt gccgaagaaa ggcccacccg
     4621 tgaaggtgag ccagtgagtt gattgctacg taattagtta gttagccctt agtgactcga
     4681 attcgcggcc gcttctagag tataaacgca gaaaggccca cccgaaggtg agccagtgtg
     4741 actctagtag agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     4801 ctgagccttt cgttttattt gatgcctggc tctagtatta ttacttgtac agctcgtcca
     4861 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     4921 cgttggggtc tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca
     4981 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     5041 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     5101 tgatatagac gttgtcgcta aagtagttgt actccagctt gtgccccagg atgttgccgt
     5161 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     5221 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     5281 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     5341 gggcgaagca ctgcacgccc caggacaggg tggtcacgag ggtgggccag ggcacgggca
     5401 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     5461 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     5521 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ctagtatttc tcctctttct
     5581 ctagtagcta gcataatacc taggactgag ctagctgtaa atactagtag cggccgctgc
     5641 aggagtcact aagggttagt tagttagatt agcagaaagt caaaagcctc cgaccggagg
     5701 cttttgacta aaacttccct tggggttatc attggggctc actcaaaggc ggtaatcaga
     5761 taaaaaaaat ccttagcttt cgctaaggat gatttctgct agagctgtca gaccaagttt
     5821 acgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     5881 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagg
     5941 gacagtaaga cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt
     6001 ctgaatgacc tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa
     6061 ctgctgaaca gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga
     6121 gaagcccgtg acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag
     6181 gcgcctgtag tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat
     6241 gtcacgaaaa agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg
     6301 atttgcccga gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta
     6361 gaggagcaaa cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg
     6421 agttatacac agggctggga tctattcttt ttatcttttt ttattctttc tttattctat
     6481 aaattataac cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag
     6541 agggtctagc agaatttaca agttttccag caaaggtcta gcagaattta cagataccca
     6601 caactcaaag gaaaaggaca tgtaattatc attgactagc ccatctcaat tggtatagtg
     6661 attaaaatca c
//
