LOCUS       sequence-488096-        3161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Temperature-Inducible promoter, including ribosome 
            binding site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3161)
  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  JOURNAL   Unpublished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 3161)
  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            "Unpublished"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_FULLPRM_cI857_PR
FEATURES             Location/Qualifiers
     source          1..3161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             814..822
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(866..884)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(867..884)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(900..916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(900..916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(1031..1619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1111..1130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1734..1836
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1837..2496
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1903..1922)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2609..3064
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2696..2715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2906..2927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3083..3100
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3084..3100
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3107..3126
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3107..3125
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcggga gtcagccaaa cgtttcttca ggccactgac tagcgataac tttccccaca
       61 acggaacaac tctcattgca tgggatcatt gggtactgtg ggtttagtgg ttgtaaaaac
      121 acctgaccgc tatccctgat cagtttcttg aaggtaaact catcaccccc aagtctggct
      181 atgcagaaat cacctggctc aacagcctgc tcagggtcaa cgagaattaa cattccgtca
      241 ggaaagcttg gcttggagcc tgttggtgcg gtcatggaat taccttcaac ctcaagccag
      301 aatgcagaat cactggcttt tttggttgtg cttacccatc tctccgcatc acctttggta
      361 aaggttctaa gctcaggtga gaacatccct gcctgaacat gagaaaaaac agggtactca
      421 tactcacttc taagtgacgg ctgcatacta accgcttcat acatctcgta gatttctctg
      481 gcgattgaag ggctaaattc ttcaacgcta actttgagaa tttttgtaag caatgcggcg
      541 ttataagcat ttaatgcatt gatgccatta aataaagcac caacgcctga ctgccccatc
      601 cccatcttgt ctgcgacaga ttcctgggat aagccaagtt catttttctt tttttcataa
      661 attgctttaa ggcgacgtgc gtcctcaagc tgctcttgtg ttaatggttt cttttttgtg
      721 ctcatacgtt aaatctatca ccgcaaggga taaatatcta acaccgtgcg tgttgactat
      781 tttacctctg gcggtgataa tggttgcatg tactaaggag gttgtaatgc gagaccactc
      841 atcgcccaat cactagtgaa ttcgttctat agtgtcacct aaataccata gagccgttgg
      901 tcatagctgt ttcctgtcca ggcatcatga cacggttatc cacagaatca ggggataacg
      961 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1081 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1141 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1201 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1261 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1321 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1381 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1441 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1501 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1561 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg tgatgatcgg
     1741 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     1801 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     1861 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     1921 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     1981 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     2041 gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     2101 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     2161 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     2221 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt gtcagccaat
     2281 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     2341 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     2401 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     2461 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     2521 ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat
     2581 tcgatgataa gctgtcaaac atgatgttac gcgccctgta gcggcgcatt aagcgcggcg
     2641 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2701 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2761 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2821 gattagggtg atggttcacg tagtgggcca gcgccctgat agacggtttt tcgccctttg
     2881 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2941 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3001 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3061 attttaacat tgtagaaacg catgtaaaac gacggccagt gaattgtaat acgactcact
     3121 atagggtgaa ttccgcttag gaccttagat attgatctca a
//
