LOCUS       40924_38768        4311 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BioBrick cloning vector pSB4C5-I52002, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4311)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Engineering BioBrick vectors from BioBrick parts
  JOURNAL   J Biol Eng 2, 5 (2008)
  PUBMED    18410688
REFERENCE   2  (bases 1 to 4311)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2008) Biological Engineering, MIT, 32 Vassar 
            Street Room 32-311, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4311)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4311)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol Eng 
            2, 5 (2008)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-FEB-2008) Biological Engineering, MIT, 32 Vassar Street Room 
            32-311, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..21
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     misc_feature    35..46
                     /note="translational stop sequence for BioBrick
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     terminator      57..88
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     primer_bind     complement(116..135)
                     /note="VR reverse verfication primer annealing site 
                     (BBa_G00102)"
     regulatory      136..176
                     /note="derived from ribosomal RNA rrnC operon, reverse 
                     orientation (BBa_B0062); similar to Escherichia coli strain
                     MG1655 accession U00096"
                     /regulatory_class="terminator"
     stem_loop       complement(149..168)
                     /label=derived from ribosomal RNA rrnC operon
                     /note="derived from ribosomal RNA rrnC operon"
     terminator      213..307
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    320..687
                     /label=par region
                     /note="par region"
     rep_origin      799..1021
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     regulatory      1033..1038
                     /gene="rep101"
                     /regulatory_class="minus_10_signal"
     regulatory      1053..1065
                     /gene="rep101"
                     /regulatory_class="ribosome_binding_site"
     regulatory      1062..1067
                     /gene="rep101"
                     /regulatory_class="ribosome_binding_site"
     CDS             1069..2016
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     promoter        2231..2334
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2335..2991
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      3005..3064
                     /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."
     primer_bind     3077..3096
                     /note="VF2 forward verification primer annealing site 
                     (BBa_G00100)"
     terminator      complement(3124..3153)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    3175..3186
                     /note="translational stop sequence for BioBrick
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     misc_feature    3200..3221
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      3262..3267
                     /gene="ccdB"
                     /regulatory_class="minus_35_signal"
     regulatory      3285..3290
                     /gene="ccdB"
                     /regulatory_class="minus_10_signal"
     CDS             3329..3631
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      3664..4252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tactagtagc ggccgctgca ggagtcacta agggttagtt agttagatta gcagaaagtc
       61 aaaagcctcc gaccggaggc ttttgactaa aacttccctt ggggttatca ttggggctca
      121 ctcaaaggcg gtaatcagat aaaaaaaatc cttagctttc gctaaggatg atttctgcta
      181 gagctgtcag accaagttta cgagctcgct tggactcctg ttgatagatc cagtaatgac
      241 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
      301 tgagaatcca agcactaggg acagtaagac gggtaagcct gttgatgata ccgctgcctt
      361 actgggtgca ttagccagtc tgaatgacct gtcacgggat aatccgaagt ggtcagactg
      421 gaaaatcaga gggcaggaac tgctgaacag caaaaagtca gatagcacca catagcagac
      481 ccgccataaa acgccctgag aagcccgtga cgggcttttc ttgtattatg ggtagtttcc
      541 ttgcatgaat ccataaaagg cgcctgtagt gccatttacc cccattcact gccagagccg
      601 tgagcgcagc gaactgaatg tcacgaaaaa gacagcgact caggtgcctg atggtcggag
      661 acaaaaggaa tattcagcga tttgcccgag cttgcgaggg tgctacttaa gcctttaggg
      721 ttttaaggtc tgttttgtag aggagcaaac agcgtttgcg acatcctttt gtaatactgc
      781 ggaactgact aaagtagtga gttatacaca gggctgggat ctattctttt tatctttttt
      841 tattctttct ttattctata aattataacc acttgaatat aaacaaaaaa aacacacaaa
      901 ggtctagcgg aatttacaga gggtctagca gaatttacaa gttttccagc aaaggtctag
      961 cagaatttac agatacccac aactcaaagg aaaaggacat gtaattatca ttgactagcc
     1021 catctcaatt ggtatagtga ttaaaatcac ctagaccaat tgagatgtat gtctgaatta
     1081 gttgttttca aagcaaatga actagcgatt agtcgctatg acttaacgga gcatgaaacc
     1141 aagctaattt tatgctgtgt ggcactactc aaccccacga ttgaaaaccc tacaaggaaa
     1201 gaacggacgg tatcgttcac ttataaccaa tacgctcaga tgatgaacat cagtagggaa
     1261 aatgcttatg gtgtattagc taaagcaacc agagagctga tgacgagaac tgtggaaatc
     1321 aggaatcctt tggttaaagg ctttgagatt ttccagtgga caaactatgc caagttctca
     1381 agcgaaaaat tagaattagt ttttagtgaa gagatattgc cttatctttt ccagttaaaa
     1441 aaattcataa aatataatct ggaacatgtt aagtcttttg aaaacaaata ctctatgagg
     1501 atttatgagt ggttattaaa agaactaaca caaaagaaaa ctcacaaggc aaatatagag
     1561 attagccttg atgaatttaa gttcatgtta atgcttgaaa ataactacca tgagtttaaa
     1621 aggcttaacc aatgggtttt gaaaccaata agtaaagatt taaacactta cagcaatatg
     1681 aaattggtgg ttgataagcg aggccgcccg actgatacgt tgattttcca agttgaacta
     1741 gatagacaaa tggatctcgt aaccgaactt gagaacaacc agataaaaat gaatggtgac
     1801 aaaataccaa caaccattac atcagattcc tacctacgta acggactaag aaaaacacta
     1861 cacgatgctt taactgcaaa aattcagctc accagttttg aggcaaaatt tttgagtgac
     1921 atgcaaagta agcatgatct caatggttcg ttctcatggc tcacgcaaaa acaacgaacc
     1981 acactagaga acatactggc taaatacgga aggatctgag gttcttatgg ctcttgtatc
     2041 tatcagtgaa gcatcaagac taacaaacaa aagtagaaca actgttcacc gttagatatc
     2101 aaagggaaaa ctgtccatat gcacagatga aaacggtgta aaaaagatag atacatcaga
     2161 gcttttacga gtttttggtg catttaaagc tgttcaccat gaacagatcg acaatgtaac
     2221 tactagaggt tgatcgggca cgtaagaggt tccaactttc accataatga aataagatca
     2281 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag
     2341 aaaaaaatca cgggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     2401 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     2461 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     2521 cttgcccgcc tgatgaacgc tcacccggag tttcgtatgg ccatgaaaga cggtgagctg
     2581 gtgatctggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     2641 tcgtccctct ggagtgaata ccacgacgat ttccggcagt ttctccacat atattcgcaa
     2701 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     2761 ttttttgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     2821 atggacaact tcttcgcccc cgttttcacg atgggcaaat attatacgca aggcgacaag
     2881 gtgctgatgc cgctggcgat ccaggttcat catgccgttt gtgatggctt ccatgtcggc
     2941 cgcatgctta atgaattaca acagtactgt gatgagtggc agggcggggc gtaataatac
     3001 tagctccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     3061 aagattactt cgcgtttgcc acctgacgtc taagaaaagg aatattcagc aatttgcccg
     3121 tgccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgctac gtaattagtt
     3181 agttagccct tagtgactcg aattcgcggc cgcttctaga gggcttacta aaagccagat
     3241 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
     3301 cccgaagtat gtcaaaaaga ggtatgctat gcagtttaag gtttacacct ataaaagaga
     3361 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     3421 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     3481 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     3541 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     3601 aaacgccatt aacctgatgt tctggggaat ataataaccc gtagaaaaga tcaaaagatc
     3661 ttcttgagat cctttttttc tgcgcgtaat ctgctacttg caaacaaaaa aaccaccgct
     3721 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3781 ctttagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3841 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3901 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3961 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4021 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4081 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4141 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4201 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4261 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca t
//
