LOCUS       sequence-138299-        2137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Secondary module (BpiI) site in pTU2-a backbone with p15A (low-copy)
            origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2137)
  AUTHORS   Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont
            PS
  TITLE     EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2016 May 2.
  PUBMED    27096716
REFERENCE   2  (bases 1 to 2137)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2016 May 2."
COMMENT     Sequence Label: pTU2S-b (p15A origin)
FEATURES             Location/Qualifiers
     source          1..2137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    42..63
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      116..173
                     /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."
     rep_origin      413..958
                     /direction=RIGHT
                     /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.
                     "
     primer_bind     859..878
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1128..1214)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1240..1334)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1355..2014)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1929..1948
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tcctataagt cttccatatg cacgcatgcg aagacaagta cgaattcgcg gccgcttcta
       61 gaggtctcac tatatctaga gacgcaccgt ctcagaaggt acagagaccc tgcagtccgg
      121 caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc
      181 gcgttatgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      241 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      301 gaaagggatc taacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
      361 taaattgcac tgaaatctag taatatttta tctgattaat aagatgatct tcttgagatc
      421 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
      481 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
      541 agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
      601 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
      661 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
      721 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
      781 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
      841 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      901 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
      961 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     1021 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     1081 tcagtgagcg aggaagcgga atatatcccc taggtctagg gcggcggatt tgtcctactc
     1141 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     1201 tttcgtttta tttgatgcct ctagtagctc gaggcttgga ttctcaccaa taaaaaacgc
     1261 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     1321 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     1381 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     1441 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     1501 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     1561 tcacccaggg attggctgac acgaaaaaca tattctcaat aaacccttta gggaaatagg
     1621 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     1681 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     1741 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     1801 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     1861 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     1921 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     1981 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     2041 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     2101 ctcttacgtg cccgatcaac tcgagtgcca cctgacg
//
