LOCUS       40924_38788        4825 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BioBrick cloning vector pSB4T5-I52001, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4825)
  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 4825)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2008) Biological Engineering, MIT, 32 Vassar 
            Street Room 32-311, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4825)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4825)
  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 
            (14-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..4825
                     /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"
     promoter        2242..2270
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             2318..3505
                     /label=TcR
                     /note="tetracycline efflux protein"
     terminator      3519..3578
                     /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     3591..3610
                     /note="VF2 forward sequencing primer annealing site 
                     (BBa_G00100)"
     terminator      complement(3638..3667)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    3689..3700
                     /note="translational stop sequence for BioBrick
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     misc_feature    3714..3735
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      3776..3781
                     /gene="ccdB"
                     /regulatory_class="minus_35_signal"
     regulatory      3799..3804
                     /gene="ccdB"
                     /regulatory_class="minus_10_signal"
     CDS             3843..4145
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      4178..4766
                     /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 tactagagga gattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt
     2281 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
     2341 cgtcattctc ggcaccgtca ccctggacgc tgtaggcata ggcttggtta tgccggtact
     2401 gccgggcctc ttgcgggata tcgtccattc cgacagtatt gccagtcact atggcgtgct
     2461 gcttgcgctc tatgcgttga tgcaatttct ttgcgcaccc gttctcggag ccctgtccga
     2521 ccgctttggc cgccgtccag tcctgctcgc ttcgctcctt ggagccacta tcgactacgc
     2581 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggcgggcat
     2641 cacgggtgcc acaggtgcgg ttgctggtgc ctatatcgcc gacatcaccg atggggaaga
     2701 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     2761 cgtggccggg ggactgttgg gtgccatctc cttgcatgca ccattccttg cggcggcggt
     2821 gctcaacggc ctcaacctcc tcctgggctg cttccttatg caggaatcgc ataagggaga
     2881 gcgccgtccg atgcccttgc gtgccttcaa tccagtcagc tccttccggt gggcgcgggg
     2941 catgactatc gtcgccgcac ttatgactgt tttctttatc atgcaactcg taggacaggt
     3001 tccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3061 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcacggg
     3121 ccccgccacc aaacgtttcg gcgagaagca ggccattatc gcgggcatgg cggccgacgc
     3181 gctgggctac gtcttgctgg cgttcgcgac gcgcggctgg atggccttcc ccattatgat
     3241 tcttctcgct tccggcggca tcggtatgcc cgcgttgcag gccatgctgt cccgccaagt
     3301 agatgacgac catcagggac agcttcaagg gtcgctcgcg gctcttacca gcctcacttc
     3361 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     3421 gttggcatgg attgtaggtg ccgcccttta ccttgtctgc ctccccgcgt tgcgtcgcgg
     3481 tgcatggagc cgggccacct cgacctaata atactagctc cggcaaaaaa acgggcaagg
     3541 tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt tgccacctga
     3601 cgtctaagaa aaggaatatt cagcaatttg cccgtgccga agaaaggccc acccgtgaag
     3661 gtgagccagt gagttgattg ctacgtaatt agttagttag cccttagtga ctcgaattcg
     3721 cggccgcttc tagagggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     3781 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg
     3841 ctatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     3901 tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac
     3961 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     4021 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     4081 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     4141 gaatataata acccgtagaa aagatcaaaa gatcttcttg agatcctttt tttctgcgcg
     4201 taatctgcta cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4261 aagagctacc aactcttttt ccgaaggtaa ctggctttag cagagcgcag ataccaaata
     4321 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4381 catacctcgc tctgctaatc ctgttaccag tagctactgc cagtagcgat aagtcgtgtc
     4441 ttaccgggtt ggattcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatctgg
     4621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4741 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4801 ccttttgctg gccttttgct cacat
//
