LOCUS       40924_2857        6644 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector p4wtVT-lacI, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  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  JOURNAL   Unpublished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 6644)
  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 6644)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6644)
  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            "Unpublished"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..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..966
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     promoter        1179..1197
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1233..1244
                     /label=BBa_B0034
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             1233..1244
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1251..1967
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
     terminator      1990..2061
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2077..2104
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      2111..2145
                     /label=PJ23101*
                     /note="PJ23101*"
                     /regulatory_class="promoter"
     RBS             2154..2165
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2181..3260
                     /label=lacI
                     /note="lac repressor"
     CDS             3261..3293
                     /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(3344..3387)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3428..3455
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3462..3482
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             3586..4443
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      4457..4516
                     /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(4576..4605)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    4652..4673
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     terminator      complement(4680..4707)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4723..4794)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4817..5533)
                     /label=mTurquoise2
                     /note="enhanced monomeric variant of CFP (Goedhart et al., 
                     2012)"
     regulatory      complement(5534..5551)
                     /label=BBa_B0034
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             5540..5551
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(5560..5594)
                     /label=pJ23101
                     /note="pJ23101"
                     /regulatory_class="promoter"
     misc_feature    5595..5615
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5651..5682
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     terminator      5807..5901
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      6393..6615
                     /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 atacgactca ctatagggaa
     1201 tacaagctac ttgttctttt tgcatactag agaaagagga gaaatactag atggtgagca
     1261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1321 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1381 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1441 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1681 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1741 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1861 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa taatactaga
     1981 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2041 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     2101 tgcgtttata tttacagcta gctcagtcct aggttttatg ctagctacta gagaaagagg
     2161 agaaatacta gatggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     2221 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     2281 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     2341 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     2401 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     2461 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     2521 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     2581 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     2641 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     2701 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     2761 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     2821 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     2881 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     2941 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     3001 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     3061 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     3121 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     3181 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3241 cccgactgga aagcgggcag gctgcaaacg acgaaaacta cgctttagta gcttaataac
     3301 tctgatagtg ctagtgtaga tctctactag agccaggcat caaataaaac gaaaggctca
     3361 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     3421 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagtag cggccgctgc
     3481 agtgatctac actagcacta tcagtgctaa atacattcaa atatctatcc gctcatgaga
     3541 caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat
     3601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3661 gaaacgctgg tgaaagtaaa agatgccgaa gatcagttgg gtgcacgtgt gggttacatc
     3721 gaactggacc tcaacagcgg taagattctt gagagttttc gccccgaaga acgtttccca
     3781 atgatgagca cttttaaagt tctgctctgt ggcgcggtat tatcccgtat tgacgccggg
     3841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3901 gtcacagaaa agcatcttac ggacggcatg acagtacgcg aattatgcag cgctgccata
     3961 accatgagtg ataacacggc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4021 cttaccgctt ttttgcacaa catgggtgat catgtaactc gccttgatcg ttgggaaccg
     4081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agctatggca
     4141 acaacgttgc gcaaactctt aactggcgaa cttcttactc tcgcttcccg gcaacaatta
     4201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcccgcgg tattattgca
     4321 gccctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     4381 gcaactatgg acgaacgtaa tcgccagatc gctgagatag gtgcctccct gattaagcat
     4441 tggtaataat actagctccg gcaaaaaaac gggcaaggtg tcaccaccct gccctttttc
     4501 tttaaaaccg aaaagattac ttcgcgtttg ccacctgacg tctaagaaaa ggaatattca
     4561 gcaatttgcc cgtgccgaag aaaggcccac ccgtgaaggt gagccagtga gttgattgct
     4621 acgtaattag ttagttagcc cttagtgact cgaattcgcg gccgcttcta gagtataaac
     4681 gcagaaaggc ccacccgaag gtgagccagt gtgactctag tagagagcgt tcaccgacaa
     4741 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     4801 ggctctagta ttattacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     4861 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     4921 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     4981 gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt
     5041 tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtcg ctaaagtagt
     5101 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     5161 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     5221 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     5281 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccccaggaca
     5341 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     5401 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     5461 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     5521 ccttgctcac catctagtat ttctcctctt tctctagtag ctagcataat acctaggact
     5581 gagctagctg taaatactag tagcggccgc tgcaggagtc actaagggtt agttagttag
     5641 attagcagaa agtcaaaagc ctccgaccgg aggcttttga ctaaaacttc ccttggggtt
     5701 atcattgggg ctcactcaaa ggcggtaatc agataaaaaa aatccttagc tttcgctaag
     5761 gatgatttct gctagagctg tcagaccaag tttacgagct cgcttggact cctgttgata
     5821 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     5881 gcgtttttta ttggtgagaa tccaagcact agggacagta agacgggtaa gcctgttgat
     5941 gataccgctg ccttactggg tgcattagcc agtctgaatg acctgtcacg ggataatccg
     6001 aagtggtcag actggaaaat cagagggcag gaactgctga acagcaaaaa gtcagatagc
     6061 accacatagc agacccgcca taaaacgccc tgagaagccc gtgacgggct tttcttgtat
     6121 tatgggtagt ttccttgcat gaatccataa aaggcgcctg tagtgccatt tacccccatt
     6181 cactgccaga gccgtgagcg cagcgaactg aatgtcacga aaaagacagc gactcaggtg
     6241 cctgatggtc ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg agggtgctac
     6301 ttaagccttt agggttttaa ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc
     6361 ttttgtaata ctgcggaact gactaaagta gtgagttata cacagggctg ggatctattc
     6421 tttttatctt tttttattct ttctttattc tataaattat aaccacttga atataaacaa
     6481 aaaaaacaca caaaggtcta gcggaattta cagagggtct agcagaattt acaagttttc
     6541 cagcaaaggt ctagcagaat ttacagatac ccacaactca aaggaaaagg acatgtaatt
     6601 atcattgact agcccatctc aattggtata gtgattaaaa tcac
//
