LOCUS       40924_38778        4509 bp DNA     circular SYN 18-DEC-2018
DEFINITION  BioBrick cloning vector pSB4K5-I52002, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4509)
  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 4509)
  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 4509)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4509)
  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..4509
                     /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"
     CDS             complement(190..1002)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(1363..2310)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     regulatory      complement(2312..2317)
                     /gene="rep101"
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(2314..2326)
                     /gene="rep101"
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(2341..2346)
                     /gene="rep101"
                     /regulatory_class="minus_10_signal"
     rep_origin      complement(2358..2580)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_feature    complement(2692..3059)
                     /label=par region
                     /note="par region"
     terminator      complement(3072..3166)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      3203..3262
                     /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     3275..3294
                     /note="VF2 forward sequencing primer annealing site 
                     (BBa_G00100)"
     terminator      complement(3322..3351)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    3373..3384
                     /note="translational stop sequence for BioBrick
                     vector-insert insulation, has stop codons in all 6 reading 
                     frames (BBa_B0042)"
     misc_feature    3398..3419
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      3460..3465
                     /gene="ccdB"
                     /regulatory_class="minus_35_signal"
     regulatory      3483..3488
                     /gene="ccdB"
                     /regulatory_class="minus_10_signal"
     CDS             3527..3829
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      3862..4450
                     /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 gtattattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
      241 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
      301 gttccaaaga atggcaaggt cctggtaacg gtctgcgatt ccgacccgtc caacatcaat
      361 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
      421 gacgactgaa tccggtgaga atggcaagag cttgtgcatt tctttccaga cttgttcaac
      481 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcatgcg
      541 tgattgcgcc tgagcaagac gaaatacacg atcgctgtta aaaggacaat tacaaacagg
      601 aatcgaatgt aaccggcgca ggaacacggc cagcgcatca acaatatttt cacctgaatc
      661 aggatattct tctaatacct ggaaggctgt tttcccagga atcgcggtgg tgagtaacca
      721 cgcatcatca ggagtacgga taaaatgctt gatggtcggg agaggcataa actccgtcag
      781 ccagttgaga cggaccatct catctgtaac atcattggca acgctacctt tgccatgttt
      841 cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg
      901 cccgacatta tcgcgagccc atttataccc atataaatca gcgtccatgt tggagtttaa
      961 gcgcggacgg gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     1021 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     1081 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     1141 gaaggatcag ctctagtagt tacattgtcg atctgttcat ggtgaacagc tttaaatgca
     1201 ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat
     1261 atggacagtt ttccctttga tatctaacgg tgaacagttg ttctactttt gtttgttagt
     1321 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     1381 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     1441 atcatgctta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     1501 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta cgtaggtagg aatctgatgt
     1561 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     1621 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     1681 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     1741 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     1801 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     1861 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     1921 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     1981 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     2041 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     2101 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     2161 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     2221 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     2281 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     2341 actataccaa ttgagatggg ctagtcaatg ataattacat gtccttttcc tttgagttgt
     2401 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     2461 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     2521 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     2581 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     2641 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     2701 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     2761 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     2821 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     2881 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     2941 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     3001 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc
     3061 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     3121 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgta
     3181 aacttggtct gacagctcta gctccggcaa aaaaacgggc aaggtgtcac caccctgccc
     3241 tttttcttta aaaccgaaaa gattacttcg cgtttgccac ctgacgtcta agaaaaggaa
     3301 tattcagcaa tttgcccgtg ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg
     3361 attgctacgt aattagttag ttagccctta gtgactcgaa ttcgcggccg cttctagagg
     3421 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     3481 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatgc agtttaaggt
     3541 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     3601 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     3661 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     3721 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     3781 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aataacccgt
     3841 agaaaagatc aaaagatctt cttgagatcc tttttttctg cgcgtaatct gctacttgca
     3901 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3961 ttttccgaag gtaactggct ttagcagagc gcagatacca aatactgtcc ttctagtgta
     4021 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4081 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4141 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4201 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4261 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4321 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4381 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4441 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4501 tgctcacat
//
