LOCUS       sequence-121842-        4563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  regulated expression of genes in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4563)
  AUTHORS   Stark MJ
  TITLE     Multicopy expression vectors carrying the lac repressor gene for 
            regulated high-level expression of genes in Escherichia coli.
  JOURNAL   Gene. 1987;51(2-3):255-67.
  PUBMED    3110013
REFERENCE   2  (bases 1 to 4563)
  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: "Gene."; 
            date: "1987"; volume: "51(2-3)"; pages: "255-67"
COMMENT     Sequence Label: pttq18
FEATURES             Location/Qualifiers
     source          1..4563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      897..1485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1386..1405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1639..1656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(1927..1932,1933..1948,1949..1955)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1963..1979
                     /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     1968..1990
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1999..2322
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLAAH
                     PPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRINSLFWRMREDFQP
                     DTD"
     misc_feature    2001..2057
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2061..2078)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2061..2077)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2070..2092)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2308..2325)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2478..2564
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2656..2683
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2798..2817)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2917..2939
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2977..2995)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3051..3128
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3129..4211
                     /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(3148..3167)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        4324..4428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4429..4497,4498..4563,1..726)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
       61 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      121 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      181 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      241 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      301 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      361 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      421 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      481 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      541 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      601 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      661 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      721 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
      781 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
      841 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      901 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      961 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1021 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1081 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1141 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1201 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1261 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1321 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1381 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1441 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1501 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1561 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1621 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1681 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gaattaattc tcatgtttga
     1741 cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     1801 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
     1861 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
     1921 gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat
     1981 ttcacacagg aaacagcgat gaattcgagc tcggtacccg gggatcctct agagtcgacc
     2041 tgcaggcatg caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2101 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2161 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     2221 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccctg
     2281 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     2341 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     2401 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     2461 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     2521 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     2581 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     2641 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     2701 ttcctgtcgt catatctaca agccatcccc ccacagatac ggtaaactag cctcgttttt
     2761 gcatcaggaa agcagggaat ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2821 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2881 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2941 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3001 aggttaatgt catgataata atggtttctt agacgtgagg ttctgtaccc gacaccatcg
     3061 aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
     3121 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
     3181 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
     3241 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
     3301 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
     3361 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
     3421 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
     3481 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     3541 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     3601 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     3661 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     3721 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     3781 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     3841 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     3901 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     3961 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     4021 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     4081 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     4141 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4201 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
     4261 ttacacttta tgcttccgac ctgcaagaac ctcacgtcag gtggcacttt tcggggaaat
     4321 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4381 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4441 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4501 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4561 atc
//
