LOCUS       sequence-413888-        3286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type 0 promoter part .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3286)
  AUTHORS   Shapiro M
  TITLE     EcoFlex MoClo Extension, Volume I
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3286)
  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            "unpublished"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: pBP-P_lacI-Ptac-lacO
FEATURES             Location/Qualifiers
     source          1..3286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(183..277)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(298..957)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     872..891
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1119..1137
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1152..1191)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(1192..2274)
                     /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="MKPVTLYDVAEYAGVSYMTVSRVVNQASHVSAKTREKVEAAMVEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIYSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2236..2255
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2359..2364,2365..2380,2381..2387)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2393..2412
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     terminator      2446..2503
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2527..2544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2698..3286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2778..2797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ctcgaggctt
      181 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
      241 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
      301 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
      361 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
      421 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
      481 gtttaaatca aaactggtga aactcaccca gggattggct gacacgaaaa acatattctc
      541 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
      601 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
      661 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
      721 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
      781 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
      841 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
      901 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
      961 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     1021 ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aactcgagtg ccacctgacg
     1081 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga
     1141 ggtctcacta ttccaaaaaa aaaggctcca aaaggagcct ttaattgtat ctcactgccc
     1201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1261 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agactggcaa
     1321 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     1381 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     1441 atcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     1501 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     1561 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     1621 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     1681 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     1741 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcctcatggg agtaaataat
     1801 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     1861 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     1921 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     1981 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2041 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     2101 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccacca tcgccgcttc
     2161 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     2221 catataagag acaccggcat actctgcgac atcgtataac gttactggtt tcatattcac
     2281 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     2341 ttcgatggcg cgccgctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga
     2401 gcgctcacaa ttgtacagag acccatgccg tctcattagc tgcagtccgg caaaaaaggg
     2461 caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc gcgttatgca
     2521 ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2701 ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaa
//
