LOCUS       sequence-487851-        3058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Temperature-dependent transcriptional repressor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3058)
  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  JOURNAL   Unpublished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 3058)
  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            "Unpublished"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_cI857
FEATURES             Location/Qualifiers
     source          1..3058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             9..722
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     promoter        complement(763..781)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(764..781)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(797..813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(797..813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(928..1516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1008..1027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1631..1733
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1734..2393
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1800..1819)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2506..2961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2593..2612)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2803..2824
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2980..2997
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2981..2997
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3004..3023
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3004..3022
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcgaat gagcacaaaa aagaaaccat taacacaaga gcagcttgag gacgcacgtc
       61 gccttaaagc aatttatgaa aaaaagaaaa atgaacttgg cttatcccag gaatctgtcg
      121 cagacaagat ggggatgggg cagtcaggcg ttggtgcttt atttaatggc atcaatgcat
      181 taaatgctta taacgccgca ttgcttacaa aaattctcaa agttagcgtt gaagaattta
      241 gcccttcaat cgccagagaa atctacgaga tgtatgaagc ggttagtatg cagccgtcac
      301 ttagaagtga gtatgagtac cctgtttttt ctcatgttca ggcagggatg ttctcacctg
      361 agcttagaac ctttaccaaa ggtgatgcgg agagatgggt aagcacaacc aaaaaagcca
      421 gtgattctgc attctggctt gaggttgaag gtaattccat gaccgcacca acaggctcca
      481 agccaagctt tcctgacgga atgttaattc tcgttgaccc tgagcaggct gttgagccag
      541 gtgatttctg catagccaga cttgggggtg atgagtttac cttcaagaaa ctgatcaggg
      601 atagcggtca ggtgttttta caaccactaa acccacagta cccaatgatc ccatgcaatg
      661 agagttgttc cgttgtgggg aaagttatcg ctagtcagtg gcctgaagaa acgtttggct
      721 gagcttcgag accactcatc gcccaatcac tagtgaattc gttctatagt gtcacctaaa
      781 taccatagag ccgttggtca tagctgtttc ctgtccaggc atcatgacac ggttatccac
      841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1621 ccttttgtga tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1681 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     1741 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1801 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     1861 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     1921 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     1981 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     2041 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     2101 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2161 ttttcgtgtc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2221 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     2281 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2341 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt
     2401 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
     2461 ggatgaatgg cagaaattcg atgataagct gtcaaacatg atgttacgcg ccctgtagcg
     2521 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2581 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2641 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2701 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccagcg ccctgataga
     2761 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2821 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2881 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2941 aaatattaac gcttacaatt ttaacattgt agaaacgcat gtaaaacgac ggccagtgaa
     3001 ttgtaatacg actcactata gggtgaattc cgcttaggac cttagatatt gatctcaa
//
