LOCUS       40924_38763        4485 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BioBrick cloning vector pSB4A5-I52002, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4485)
  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 4485)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2008) Biological Engineering, MIT, 32 Vassar 
            Street Room 32-311, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4485)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4485)
  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 
            (12-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..4485
                     /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 sequencing 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"
     regulatory      2259..2264
                     /gene="bla"
                     /regulatory_class="minus_35_signal"
     regulatory      2285..2290
                     /gene="bla"
                     /regulatory_class="minus_10_signal"
     regulatory      2299..2303
                     /gene="bla"
                     /regulatory_class="ribosome_binding_site"
     CDS             2308..3165
                     /label=AmpR
                     /note="beta-lactamase"
     terminator      3179..3238
                     /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     3251..3270
                     /note="VF2 forward sequencing primer annealing site 
                     (BBa_G00100)"
     terminator      complement(3298..3327)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    3349..3360
                     /note="translational stop sequence for BioBrick
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     misc_feature    3374..3395
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      3436..3441
                     /gene="ccdB"
                     /regulatory_class="minus_35_signal"
     regulatory      3459..3464
                     /gene="ccdB"
                     /regulatory_class="minus_10_signal"
     CDS             3503..3805
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      3838..4426
                     /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 tactagagct aaatacattc aaatatctat ccgctcatga gacaataacc ctgataaatg
     2281 cttcaataat attgaaaaag gaagaatatg agtattcaac atttccgtgt cgcccttatt
     2341 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2401 aaagatgccg aagatcagtt gggtgcacgt gtgggttaca tcgaactgga cctcaacagc
     2461 ggtaagattc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa
     2521 gttctgctct gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2581 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2641 acggacggca tgacagtacg cgaattatgc agcgctgcca taaccatgag tgataacacg
     2701 gcggccaact tacttctgac aacgatcgga ggaccgaagg agcttaccgc ttttttgcac
     2761 aacatgggtg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2821 ccaaacgacg agcgtgacac cacgatgcct gtagctatgg caacaacgtt gcgcaaactc
     2881 ttaactggcg aacttcttac tctcgcttcc cggcaacaat taatagactg gatggaggcg
     2941 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3001 aaatctggag ccggtgagcg tgggtcccgc ggtattattg cagccctggg gccagatggt
     3061 aagccctccc gtatcgtagt tatctacacg acggggagcc aggcaactat ggacgaacgt
     3121 aatcgccaga tcgctgagat aggtgcctcc ctgattaagc attggtaata atactagctc
     3181 cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt
     3241 acttcgcgtt tgccacctga cgtctaagaa aaggaatatt cagcaatttg cccgtgccga
     3301 agaaaggccc acccgtgaag gtgagccagt gagttgattg ctacgtaatt agttagttag
     3361 cccttagtga ctcgaattcg cggccgcttc tagagggctt actaaaagcc agataacagt
     3421 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     3481 gtatgtcaaa aagaggtatg ctatgcagtt taaggtttac acctataaaa gagagagccg
     3541 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
     3601 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     3661 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     3721 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     3781 cattaacctg atgttctggg gaatataata acccgtagaa aagatcaaaa gatcttcttg
     3841 agatcctttt tttctgcgcg taatctgcta cttgcaaaca aaaaaaccac cgctaccagc
     3901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggctttag
     3961 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4321 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4381 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4441 ggccttttta cggttcctgg ccttttgctg gccttttgct cacat
//
