LOCUS       sequence-141928-        1991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Secondary module (BpiI) site in pTU2-a backbone with colE1 
            (medium-copy) origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1991)
  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 1991)
  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 (colE1 origin)
FEATURES             Location/Qualifiers
     source          1..1991
                     /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."
     primer_bind     complement(197..214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      418..1006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     907..926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1094..1188)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1209..1868)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1783..1802
                     /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 tggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
      361 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
      421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
      481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
      541 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
      601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
      661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
      721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
      781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
      841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     1021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtatt tagtctagta
     1081 gctcgaggct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     1141 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     1201 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     1261 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     1321 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     1381 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgacacgaaa
     1441 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     1501 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     1561 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     1621 accagctcac cgtctttcat tgccatacga aattccggat gagcattcat caggcgggca
     1681 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     1741 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     1801 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     1861 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
     1921 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgcccgat caactcgagt
     1981 gccacctgac g
//
