LOCUS       sequence-138340-        3140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pTU2-A backbone, with RFP negative selection and colE1 (medium-copy)
            origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3140)
  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 3140)
  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 (colE1 origin)
FEATURES             Location/Qualifiers
     source          1..3140
                     /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."
     primer_bind     complement(1346..1363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1567..2155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2056..2075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2243..2337)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2358..3017)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2932..2951
                     /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 ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1501 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1561 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1621 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1681 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     1741 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1801 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1861 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1921 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1981 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2041 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2101 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2161 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtattt
     2221 agtctagtag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     2281 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     2341 gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     2401 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     2461 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     2521 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     2581 gacacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     2641 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     2701 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     2761 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     2821 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     2881 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     2941 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     3001 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     3061 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc
     3121 aactcgagtg ccacctgacg
//
