LOCUS       sequence-178382-        4348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT LacI polymorphism with T at position 109.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4348)
  AUTHORS   Meinhardt S, Manley MW Jr, Becker NA, Hessman JA, Maher LJ 3rd, 
            Swint-Kruse L
  TITLE     Novel insights from hybrid LacI/GalR proteins: family-wide 
            functional attributes and biologically significant variation in 
            transcription repression.
  JOURNAL   Nucleic Acids Res. 2012 Nov;40(21):11139-54. doi: 
            10.1093/nar/gks806. Epub 2012 Sep 10.
  PUBMED    22965134
REFERENCE   2  (bases 1 to 4348)
  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"; doi: 
            "10.1093/nar/gks806"; journalName: "Nucleic Acids Res"; date: 
            "2012-11"; volume: "40"; issue: "21"; pages: "11139-54"
COMMENT     Sequence Label: LacI/109-pHG165c
FEATURES             Location/Qualifiers
     source          1..4348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             15..1097
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(34..53)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1634..1825
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1819..1841)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1927..2067
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1982..2001
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2082..2099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2253..2841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2333..2352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3012..3803,3804..3872))
                     /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"
     primer_bind     3635..3654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3873..3977)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4045..4063
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4216..4238
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4230..4247
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4231..4247
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
       61 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
      121 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
      181 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
      241 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
      301 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaaac ggcggtgcac aatcttctcg
      361 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
      421 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
      481 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
      541 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
      601 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
      661 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
      721 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
      781 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
      841 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
      901 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
      961 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     1021 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     1081 tggaaagcgg gcagtgatga gaattcgtaa taatatagct ctagattaat tgcgcccgtt
     1141 aacggcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     1201 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     1261 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     1321 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     1381 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     1441 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     1501 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     1561 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     1621 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     1681 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     1741 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     1801 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     1861 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     1921 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     1981 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     2041 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     2101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2161 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2221 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2281 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2461 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2521 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2581 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2641 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2701 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2761 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2821 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2881 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2941 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3001 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3061 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3121 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3181 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3241 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3301 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3361 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3421 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3481 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3541 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3601 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3661 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3721 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3781 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3841 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3901 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3961 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     4021 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     4081 tccgttaacg gcattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4141 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4201 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttggct
     4261 gcaggtcgac ggatccccgg gaattccacc atcgaatggt gcaaaacctt tcgcggtatg
     4321 gcatgatagc gcccggaaga gagtcaat
//
