LOCUS       sequence-138311-        3286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTU2-A backbone, with RFP negative selection and p15A (low-copy) 
            origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3286)
  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 3286)
  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: pTU2-A-RFP (p15A origin)
FEATURES             Location/Qualifiers
     source          1..3286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..57
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(61..104)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    107..128
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    262..283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(298..303,304..321,322..328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    336..352
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             370..381
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             388..1065
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1110..1181
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1197..1224
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1265..1322
                     /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      1562..2107
                     /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     2008..2027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2277..2363)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2389..2483)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2504..3163)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3078..3097
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga
       61 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
      121 ttctagaggt ctcactatat ctagagacgt ctagacatat gcaatacgca aaccgcctct
      181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      241 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      301 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      361 atactagaga aagaggagaa atactagatg gcttcctccg aagacgttat caaagagttc
      421 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt
      481 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt
      541 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct
      601 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc
      661 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc
      721 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc
      781 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac
      841 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt
      901 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt
      961 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt
     1021 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg ctgatagtgc
     1081 tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1141 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     1201 caccttcggg tgggcctttc tgcgtttata gcatgccgtc tcattaggta cagagaccct
     1261 gcagtccggc aaaaaagggc aaggtgtcac caccctgccc tttttcttta aaaccgaaaa
     1321 gattacttcg cgttatgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1441 ataacgcagg aaagggatct aacaacttat atcgtatggg gctgacttca ggtgctacat
     1501 ttgaagagat aaattgcact gaaatctagt aatattttat ctgattaata agatgatctt
     1561 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     1621 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     1681 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     1741 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     1801 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     1861 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     1921 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     1981 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2041 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     2101 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     2161 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     2221 cgtagcgagt cagtgagcga ggaagcggaa tatatcccct aggtctaggg cggcggattt
     2281 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     2341 gactgagcct ttcgttttat ttgatgcctc tagtagctcg aggcttggat tctcaccaat
     2401 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     2461 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     2521 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     2581 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     2641 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     2701 tggtgaaact cacccaggga ttggctgaca cgaaaaacat attctcaata aaccctttag
     2761 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     2821 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     2881 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     2941 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     3001 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     3061 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     3121 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     3181 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     3241 agttggaacc tcttacgtgc ccgatcaact cgagtgccac ctgacg
//
