LOCUS       sequence-488090-        3628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 IPTG/Lactose-Inducible promoter, including ribosome 
            binding site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3628)
  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  JOURNAL   Unpublished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 3628)
  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            "Unpublished"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_lacI_Plac
FEATURES             Location/Qualifiers
     source          1..3628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             88..1170
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(107..126)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1183..1204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1219..1224,1225..1242,1243..1249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1257..1273
                     /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)."
     primer_bind     1262..1284
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        complement(1333..1351)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1334..1351)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1367..1383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1367..1383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(1498..2086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1578..1597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2201..2303
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2304..2963
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2370..2389)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3076..3531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3163..3182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3373..3394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3550..3567
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3551..3567
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3574..3593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3574..3592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcggga gcaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
       61 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
      121 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
      181 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
      241 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
      301 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
      361 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
      421 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
      481 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
      541 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
      601 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
      661 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
      721 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
      781 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
      841 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
      901 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
      961 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     1021 gcggtgaagg gcaatcagct gttgcccgta tcactggtga aaagaaaaac caccctggcg
     1081 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1141 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     1201 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     1261 gagcggataa caatttcaca caggaaacag ctaatgcgag accactcatc gcccaatcac
     1321 tagtgaattc gttctatagt gtcacctaaa taccatagag ccgttggtca tagctgtttc
     1381 ctgtccaggc atcatgacac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1441 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1501 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1561 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1621 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1681 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1741 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1801 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1861 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1921 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1981 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2041 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2101 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2161 gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac gtaagaggtt
     2221 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     2281 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     2341 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     2401 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     2461 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     2521 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     2581 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2641 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2701 atttccctaa agggtttatt gagaatatgt ttttcgtgtc agccaatccc tgggtgagtt
     2761 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2821 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2881 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2941 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     3001 gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg atgataagct
     3061 gtcaaacatg atgttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3121 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3181 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3241 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3301 gttcacgtag tgggccagcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3361 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3421 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3481 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt ttaacattgt
     3541 agaaacgcat gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggtgaattc
     3601 cgcttaggac cttagatatt gatctcaa
//
