LOCUS       40924_38748        4026 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BioBrick cloning vector pSB3K5-I52002, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4026)
  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 4026)
  AUTHORS   Shetty RP, Endy D, Knight TF Jr.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-FEB-2008) Biological Engineering, MIT, 32 Vassar 
            Street Room 32-311, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4026)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4026)
  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 
            (15-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..4026
                     /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"
     rep_origin      599..1143
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             1894..2706
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      2720..2779
                     /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     2792..2811
                     /note="VF2 forward sequencing primer annealing site 
                     (BBa_G00100)"
     terminator      complement(2839..2868)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    2890..2901
                     /note="translational stop sequence for BioBrick
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     misc_feature    2915..2936
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      2977..2982
                     /gene="ccdB"
                     /regulatory_class="minus_35_signal"
     regulatory      3000..3005
                     /gene="ccdB"
                     /regulatory_class="minus_10_signal"
     CDS             3044..3346
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      3379..3967
                     /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 gagatggaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      241 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      301 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      361 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      421 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      481 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      541 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cttaataaga tgatcttctt
      601 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
      661 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
      721 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
      781 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
      841 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
      901 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
      961 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     1021 agagcgcacg agggagccgc caggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1081 cgccaccact gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg
     1141 aaaaacggct ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca
     1201 ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta
     1261 gcgagtcagt gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg
     1321 gtgcagcctt ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac
     1381 atagtaagcc agtatacact ccgctagcgc tgaggtctgc ctcgtgaaga aggtgttgct
     1441 gactcatacc aggcctgaat cgccccatca tccagccaga aagtgaggga gccacggttg
     1501 atgagagctt tgttgtaggt ggaccagttg gtgattttga acttttgctt tgccacggaa
     1561 cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcgattt
     1621 attcaacaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1681 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgtttac
     1741 tagagctgat ccttcaactc agcaaaagtt cgatttattc aacaaagcca cgttgtgtct
     1801 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     1861 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     1921 tgctcccgtc cgcgcttaaa ctccaacatg gacgctgatt tatatgggta taaatgggct
     1981 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2041 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2101 gtccgtctca actggctgac ggagtttatg cctctcccga ccatcaagca ttttatccgt
     2161 actcctgatg atgcgtggtt actcaccacc gcgattcctg ggaaaacagc cttccaggta
     2221 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggccgt gttcctgcgc
     2281 cggttacatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgtgt atttcgtctt
     2341 gctcaggcgc aatcacgcat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2401 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc acaagctctt gccattctca
     2461 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2521 aaattaatag gttgtattga tgttggacgg gtcggaatcg cagaccgtta ccaggacctt
     2581 gccattcttt ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2641 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2701 tttttctaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt
     2761 ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga aaaggaatat
     2821 tcagcaattt gcccgtgccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt
     2881 gctacgtaat tagttagtta gcccttagtg actcgaattc gcggccgctt ctagagggct
     2941 tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt ataagaatat
     3001 atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgcagt ttaaggttta
     3061 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     3121 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     3181 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     3241 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     3301 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataat aacccgtaga
     3361 aaagatcaaa agatcttctt gagatccttt ttttctgcgc gtaatctgct acttgcaaac
     3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3481 tccgaaggta actggcttta gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4021 tcacat
//
