LOCUS       sequence-488099-        2432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 T7 RNA polymerase and IPTG/Lactose-dependent 
            promoter, including ribosome binding site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2432)
  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  JOURNAL   Unpublished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 2432)
  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            "Unpublished"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_PT7lac
FEATURES             Location/Qualifiers
     source          1..2432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..30
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    31..55
                     /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             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(137..155)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(138..155)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(171..187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(171..187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(302..890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(382..401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1005..1107
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1108..1767
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1174..1193)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1880..2335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1967..1986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2177..2198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2354..2371
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2355..2371
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2378..2396
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcggga gtaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
       61 agaaataatt ttgtttaact ttaagaagga gatataaatg cgagaccact catcgcccaa
      121 tcactagtga attcgttcta tagtgtcacc taaataccat agagccgttg gtcatagctg
      181 tttcctgtcc aggcatcatg acacggttat ccacagaatc aggggataac gcaggaaaga
      241 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      301 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      361 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      421 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      481 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      541 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      601 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      661 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      721 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      781 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      841 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      901 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      961 tcatgagatt atcaaaaagg atcttcacct agatcctttt gtgatgatcg gcacgtaaga
     1021 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     1081 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1141 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1201 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1261 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1321 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1381 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1441 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1501 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tgtcagccaa tccctgggtg
     1561 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1621 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1681 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1741 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     1801 cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgatgata
     1861 agctgtcaaa catgatgtta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1921 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1981 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2041 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2101 gatggttcac gtagtgggcc agcgccctga tagacggttt ttcgcccttt gacgttggag
     2161 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2221 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2281 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aattttaaca
     2341 ttgtagaaac gcatgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggtga
     2401 attccgctta ggaccttaga tattgatctc aa
//
