LOCUS       sequence-487850-        2968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Tetracycline/Anhydrotetracycline-dependent 
            transcriptional repressor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2968)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2968)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pUPD3_CDS_tetR
FEATURES             Location/Qualifiers
     source          1..2968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(12..635)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     543..562
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(673..691)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(674..691)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(707..723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(707..723)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(838..1426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(918..937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1541..1643
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1644..2303
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1710..1729)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2416..2871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2503..2522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2713..2734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2890..2907
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2891..2907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2914..2933
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2914..2932
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcgaag cttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc
       61 aattcaaggc cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct
      121 tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact
      181 tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca
      241 tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga
      301 gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga
      361 cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc
      421 ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg
      481 agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc
      541 tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg
      601 ctgttaatca ctttactttt atctaatcta gacattcgag accactcatc gcccaatcac
      661 tagtgaattc gttctatagt gtcacctaaa taccatagag ccgttggtca tagctgtttc
      721 ctgtccaggc atcatgacac ggttatccac agaatcaggg gataacgcag gaaagaacat
      781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1021 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1261 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1501 gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac gtaagaggtt
     1561 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1621 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     1681 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1741 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1801 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     1861 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     1921 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     1981 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2041 atttccctaa agggtttatt gagaatatgt ttttcgtgtc agccaatccc tgggtgagtt
     2101 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2161 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2221 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2281 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     2341 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg atgataagct
     2401 gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2461 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2521 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2581 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2641 gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2701 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2761 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2821 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt ttaacattgt
     2881 agaaacgcat gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggtgaattc
     2941 cgcttaggac cttagatatt gatctcaa
//
