LOCUS       sequence-487848-        3427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 IPTG/Lactose-dependent transcriptional repressor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3427)
  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  JOURNAL   Unpublished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 3427)
  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            "Unpublished"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_lacI
FEATURES             Location/Qualifiers
     source          1..3427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(12..1094)
                     /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     1056..1075
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1150)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1133..1150)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1166..1182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1166..1182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(1297..1885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1377..1396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2000..2102
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2103..2762
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2169..2188)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2875..3330
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2962..2981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3172..3193
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3349..3366
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3350..3366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3373..3392
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3373..3391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcgaag ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
       61 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
      121 ttcaccagtg atacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
      181 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
      241 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
      301 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
      361 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
      421 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
      481 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
      541 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
      601 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
      661 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
      721 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
      781 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
      841 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
      901 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
      961 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     1021 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     1081 gttactggtt tcattcgaga ccactcatcg cccaatcact agtgaattcg ttctatagtg
     1141 tcacctaaat accatagagc cgttggtcat agctgtttcc tgtccaggca tcatgacacg
     1201 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1261 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1321 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1381 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1441 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1501 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1561 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1621 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1681 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1741 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1921 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1981 cacctagatc cttttgtgat gatcggcacg taagaggttc caactttcac cataatgaaa
     2041 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     2101 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     2161 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     2221 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2281 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     2341 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     2401 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     2461 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2521 agaatatgtt tttcgtgtca gccaatccct gggtgagttt caccagtttt gatttaaacg
     2581 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2641 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2701 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2761 aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg
     2821 ataataagcg gatgaatggc agaaattcga tgataagctg tcaaacatga tgttacgcgc
     2881 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2941 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3001 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3061 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccagcgc
     3121 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3181 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3241 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3301 attttaacaa aatattaacg cttacaattt taacattgta gaaacgcatg taaaacgacg
     3361 gccagtgaat tgtaatacga ctcactatag ggtgaattcc gcttaggacc ttagatattg
     3421 atctcaa
//
