LOCUS       V002615                10726 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002615
VERSION     V002615
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10726)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Site-specific chromosomal integration of large synthetic constructs
  JOURNAL   Nucleic Acids Res. 38 (6), E92 (2010)
   PUBMED   20047970
REFERENCE   2  (bases 1 to 10726)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2009) Molecular Biology, Princeton University,
            Washington Rd., Princeton, NJ 08544, USA
REFERENCE   3  (bases 1 to 10726)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10726)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
            Acids Res."; date: "2010"; volume: "38"; issue: "6"; pages: "E92"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (17-DEC-2009) Molecular Biology, Princeton University, Washington
            Rd., Princeton, NJ 08544, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(147..1022)
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     promoter        1049..1333
                     /label="araBAD promoter"
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite
                     direction (Guzman et al., 1995)"
     RBS             1360..1368
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1383..2075
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     CDS             2663..3451
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     CDS             complement(3781..4458)
                     /label="Exo"
                     /note="5' to 3' double-stranded DNA exonuclease in the
                     lambda Red system"
     CDS             complement(4458..5240)
                     /label="Beta"
                     /note="single-stranded DNA binding recombinase in the
                     lambda Red system"
     CDS             complement(5249..5662)
                     /label="Gam"
                     /note="inhibitor of the host RecBCD nuclease in the lambda
                     Red system"
     regulatory      complement(5687..5714)
                     /label="Plac"
                     /note="Plac"
                     /regulatory_class="promoter"
     protein_bind    5696..5712
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by 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)."
     promoter        complement(5720..5750)
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             complement(5879..6937)
                     /label="RecA"
                     /note="DNA repair protein from E. coli"
     CDS             complement(7429..8508)
                     /label="lacI"
                     /note="lac repressor"
     regulatory      complement(8542..8574)
                     /label="PlacI"
                     /note="PlacI"
                     /regulatory_class="promoter"
     CDS             complement(8697..9644)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     rep_origin      complement(9692..9914)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ctgctcgcgc aggctgggtg ccaagctctc gggtaacatc aaggcccgat ccttggagcc
       61 cttgccctcc cgcacgatga tcgtgccgtg atcgaaatcc agatccttga cccgcagttg
      121 caaaccctca ctgatccgca tgcttatgac aacttgacgg ctacatcatt cactttttct
      181 tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag
      241 aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg
      301 gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta
      361 atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt
      421 gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc
      481 tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc
      541 agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc
      601 ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg
      661 cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg
      721 cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag
      781 tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt
      841 ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt
      901 cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc
      961 gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc
     1021 atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat
     1081 tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt
     1141 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc
     1201 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc
     1261 atagcatttt tatccataag attagcggat cctacctgac gctttttatc gcaactctct
     1321 actgtttctc catacccgtt tttttgggaa ttcgagctct aaggaggtta taaaaaatgc
     1381 atcaaaaaaa ccaggtaatg aacctgggtc cgaactctaa actgctgaaa gaatacaaat
     1441 cccagctgat cgaactgaac atcgaacagt tcgaagcagg tatcggtctg atcctgggtg
     1501 atgcttacat ccgttctcgt gatgaaggta aaacctactg tatgcagttc gagtggaaaa
     1561 acaaagcata catggaccac gtatgtctgc tgtacgatca gtgggtactg tccccgccgc
     1621 acaaaaaaca acgtgttaac cacctgggta acctggtaat cacctggggc gcccagactt
     1681 tcaaacacca agctttcaac aaactggcta acctgttcat cgttaacaac aaaaaaacca
     1741 tcccgaacaa cctggttgaa aactacctga ccccgatgtc tctggcatac tggttcatgg
     1801 atgatggtgg taaatgggat tacaacaaaa actctaccaa caaatcgatc gtactgaaca
     1861 cccagtcttt cactttcgaa gaagtagaat acctggttaa gggtctgcgt aacaaattcc
     1921 aactgaactg ttacgtaaaa atcaacaaaa acaaaccgat catctacatc gattctatgt
     1981 cttacctgat cttctacaac ctgatcaaac cgtacctgat cccgcagatg atgtacaaac
     2041 tgccgaacac tatctcctcc gaaactttcc tgaaataagc atgcccgttc catacagaag
     2101 ctgggcgaac aaacgatgct cgccttccag aaaaccgagg atgcgaacca cttcatccgg
     2161 ggtcagcacc accggcaagc gccgcgacgg ccgaggtctt ccgatctcct gaagccaggg
     2221 cagatccgtg cacagcacct tgccgtagaa gaacagcaag gccgccaatg cctgacgatg
     2281 cgtggagacc gaaaccttgc gctcgttcgc cagccaggac agaaatgcct cgacttcgct
     2341 gctgcccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     2401 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     2461 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     2521 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     2581 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     2641 ccctaaaaca aagttaaaca tcatgaggga agcggtgatc gccgaagtat cgactcaact
     2701 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     2761 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     2821 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     2881 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     2941 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     3001 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     3061 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     3121 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     3181 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     3241 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     3301 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     3361 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     3421 gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta acaattcgtt caagccgacg
     3481 ccgcttcgcg gcgcggctta actcaagcgt tagatgcact aagcacataa ttgctcacag
     3541 ccaaactatc aggtcaagtc tgcttttatt atttttaagc gtgcataata agccctacac
     3601 aaattgggag atatatcatg aaaggctggc tttttcttgt tatcgcaata gttggcgaag
     3661 taatcgcaac atccgcatta aaatctagcg agggctttac taagctcctg cagagatctg
     3721 aattccctag agagacgaaa gtgattgcgc ctacccggat attatcgtga ggatgcgtca
     3781 tcgccattgc tccccaaata caaaaccaat ttcagccagt gcctcgtcca ttttttcgat
     3841 gaactccggc acgatctcgt caaaactcgc catgtacttt tcatcccgct caatcacgac
     3901 ataatgcagg ccttcacgct tcatacgcgg gtcatagttg gcaaagtacc aggcattttt
     3961 tcgcgtcacc cacatgctgt actgcacctg ggccatgtaa gctgacttta tggcctcgaa
     4021 accaccgagc cggaacttca tgaaatcccg ggaggtaaac gggcatttca gttcaaggcc
     4081 gttgccgtca ctgcataaac catcgggaga gcaggcggta cgcatacttt cgtcgcgata
     4141 gatgatcggg gattcagtaa cattcacgcc ggaagtgaat tcaaacaggg ttctggcgtc
     4201 gttctcgtac tgttttcccc aggccagtgc tttagcgtta acttccggag ccacaccggt
     4261 gcaaacctca gcaagcaggg tgtggaagta ggacattttc atgtcaggcc acttctttcc
     4321 ggagcggggt tttgctatca cgttgtgaac ttctgaagcg gtgatgacgc cgagccgtaa
     4381 tttgtgccac gcatcatccc cctgttcgac agctctcaca tcgatcccgg tacgctgcag
     4441 gataatgtcc ggtgtcatgc tgccaccttc tgctctgcgg ctttctgttt caggaatcca
     4501 agagctttta ctgcttcggc ctgtgtcagt tctgacgatg cacgaatgtc gcggcgaaat
     4561 atctgggaac agagcggcaa taagtcgtca tcccatgttt tatccagggc gatcagcaga
     4621 gtgttaatct cctgcatggt ttcatcgtta accggagtga tgtcgcgttc cggctgacgt
     4681 tctgcagtgt atgcagtatt ttcgacaatg cgctcggctt catccttgtc atagatacca
     4741 gcaaatccga aggccagacg ggcacactga atcatggctt tatgacgtaa catccgtttg
     4801 ggatgcgact gccacggccc cgtgatttct ctgccttcgc gagttttgaa tggttcgcgg
     4861 cggcattcat ccatccattc ggtaacgcag atcggatgat tacggtcctt gcggtaaatc
     4921 cggcatgtac aggattcatt gtcctgctca aagtccatgc catcaaactg ctggttttca
     4981 ttgatgatgc gggaccagcc atcaacgccc accaccggaa cgatgccatt ctgcttatca
     5041 ggaaaggcgt aaatttcttt cgtccacgga ttaaggccgt actggttggc aacgatcagt
     5101 aatgcgatga actgcgcatc gctggcatca cctttaaatg ccgtctggcg aagagtggtg
     5161 atcagttcct gtgggtcgac agaatccatg ccgacacgtt cagccagctt cccagccagc
     5221 gttgcgagtg cagtactcat tcgttttata cctctgaatc aatatcaacc tggtggtgag
     5281 caatggtttc aaccatgtac cggatgtgtt ctgccatgcg ctcctgaaac tcaacatcgt
     5341 catcaaacgc acgggtaatg gattttttgc tggccccgtg gcgttgcaaa tgatcgatgc
     5401 atagcgattc aaacaggtgc tggggcaggc ctttttccat gtcgtctgcc agttctgcct
     5461 ctttctcttc acgggcgagc tgctggtagt gacgcgccca gctctgagcc tcaagacgat
     5521 cctgaatgta ataagcgttc atggctgaac tcctgaaata gctgtgaaaa tatcgcccgc
     5581 gaaatgccgg gctgattagg aaaacaggaa agggggttag tgaatgcttt tgcttgatct
     5641 cagtttcagt attaatatcc attttttata agcgtcgact gtttcctgtg tgaaattgtt
     5701 atccgctcac aattccacac attatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5761 cctaatgagt gagaattcgg atctcgacgg gatgttgatt ctgtcatggc atatccttac
     5821 aacttaaaaa agcaaaaggg ccgcagatgc gacccttgtg tatcaaacaa gacgattaaa
     5881 aatcttcgtt agtttctgct acgccttcgc tatcatctac agagaaatcc ggcgttgagt
     5941 tcgggttgct cagcagcaac tcacgtactt tcttctcgat ctctttcgcg gtttccgggt
     6001 tatctttcag ccaggcagtc gcattcgctt taccctgacc gatcttctca cctttgtagc
     6061 tgtaccacgc gcctgctttc tcgatcagct tctcttttac gcccaggtca accagttcgc
     6121 cgtagaagtt gataccttcg ccgtagagga tctggaattc agcctgttta aacggcgcag
     6181 cgattttgtt cttcaccact ttcacgcggg tttcgctacc caccacgttt tcgccctctt
     6241 tcaccgcgcc gatacgacgg atgtcgagac gaacagaggc gtagaatttc agcgcgttac
     6301 caccggtagt ggtttccggg ttaccgaaca tcacaccaat tttcatacgg atctggttga
     6361 tgaagatcag cagcgtgttg gactgcttca ggttacccgc cagcttacgc atcgcctggc
     6421 tcatcatacg tgccgcaagg cccatgtgag agtcgccgat ttcgccttcg atttccgctt
     6481 tcggcgtcag tgccgccacg gagtcaacga cgataacgtc tactgcgcca gaacgcgcca
     6541 gggcgtcaca gatttccagt gcctgctcgc cggtgtccgg ctgggagcac agcaggttgt
     6601 cgatatcgac gcccagttta cgtgcgtaga ttgggtccag cgcgtgttca gcatcgataa
     6661 acgcacaggt tttaccttca cgctgcgctg cggcgatcac ctgcagcgtc agcgtggttt
     6721 taccggaaga ttccggtccg tagatttcga cgatacggcc catcggcaga ccacctgccc
     6781 caagcgcgat atccagtgaa agcgaaccgg tagagatggt ttccacatcc atggaacggt
     6841 cttcacccag gcgcatgatg gagcctttac caaattgttt ctcaatctgg cccagtgctg
     6901 ccgccaacgc tttctgtttg ttttcgtcga tagccatttt tactcctgtc atgccgggta
     6961 ataccggata gtcaatatgt tctgttgaag caattatact gtatgctcat acagtatcaa
     7021 gtgttttgta gaaattgttg ccacaaggtc tgcaatgcat acgcagtagc ctgacgacgc
     7081 accgcatcac ggtcgccgct gaagcatccc gccgggtaat gccttcaccg cgggcagtgg
     7141 caaaagcaaa ccagacggtg ccgacaggct tctctagatc cgcctacctt tcacgagttg
     7201 cgcagtttgt ctgcaagact ctatgagaag cagataagcg ataagtttgc tcaacatctt
     7261 ctcgggcata agtcggacac catggcatca cagtatcgtg atgacagagg cagggagtgg
     7321 gacaaaattg aaatcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac
     7381 aaattgataa gcaatgccaa gcttggcact ggctgatcag ctagctcact gcccgctttc
     7441 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7501 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     7561 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     7621 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     7681 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     7741 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     7801 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     7861 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     7921 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     7981 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     8041 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     8101 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     8161 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     8221 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     8281 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     8341 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     8401 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     8461 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     8521 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc accattcgat
     8581 gctagcccat gggtatggac agttttccct ttgatatgta acggtgaaca gttgttctac
     8641 ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc
     8701 cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag
     8761 aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg
     8821 gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttacgtagg
     8881 taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc
     8941 tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca
     9001 gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta
     9061 cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc
     9121 attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt
     9181 tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac
     9241 ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc
     9301 tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt tgtccactgg
     9361 aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct
     9421 ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat catctgagcg
     9481 tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg
     9541 ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga
     9601 ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct
     9661 aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt
     9721 ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat
     9781 tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa
     9841 gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc
     9901 cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg
     9961 ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca
    10021 agctcggttg cggccgcaat cgggcaaatc gctgaatatt ccttttgtct ccgaccatca
    10081 ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag
    10141 tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca
    10201 taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg
    10261 gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac
    10321 ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta
    10381 tcatcaacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    10441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    10501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    10561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    10621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    10681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccg
//
