LOCUS       sequence-178197-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:AscG transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4683)
  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 4683)
  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: LLhA-pHG165a
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     757..774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    789..929
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(855..874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1015..1037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1031..1222)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    1822..1843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1858..1863,1864..1881,1882..1888)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     1918..1934
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1918..1934
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1951..2028
                     /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."
     primer_bind     complement(2048..2067)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3292..3309)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3292..3308)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3301..3323)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3476..3494)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3562..3666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3667..3735,3736..4527)
                     /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     complement(3885..3904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      721 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      781 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
      841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
      901 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
      961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     1021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     1081 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     1141 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     1201 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     1261 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     1321 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     1381 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     1441 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     1501 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     1561 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     1621 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     1681 gcacccgtgg ccaggaccca acgctgcccg ccgttaacgg ccaatacgca aaccgcctct
     1741 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     1801 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1861 acactttatg cttccggctc gtatgttgtg tggcgtggta ctcataacaa tttcacacag
     1921 gaaacagcta tgaccatgat tacgaattcc gacaccatcg aatggtgcaa aacctttcgc
     1981 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     2041 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     2101 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     2161 ctcaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagag ccagacgctg
     2221 gggctggtag tgaccaacac gctttaccac ggcatttatt ttagtgaatt actctttcat
     2281 gccgcgcgaa tggcggaaga gaaagggcgg cagttgctat tggcagatgg taaacacagc
     2341 gcagaagaag agcgccaggc gattcagtat ctgctggatc tgcgctgcga cgcgatcatg
     2401 atttacccgc gttttttaag cgtggatgag atcgatgaca tcattgacgc gcacagtcag
     2461 ccgataatgg tgcttaatcg ccgcctgcgc aaaaacagca gccatagcgt ctggtgcgat
     2521 cataaacaga ccagttttaa cgccgtggca gagttgataa acgccgggca tcaggagatt
     2581 gctttcctta ccggctcgat ggattccccc accagcattg aacgtcttgc cgggtataaa
     2641 gacgcgctgg cgcagcatgg tattgcgctc aatgaaaaac ttatcgctaa cggtaaatgg
     2701 acgcctgcca gcggggccga aggggtagaa atgttgctcg aacgtggggc taaatttagc
     2761 gcgttagttg ccagtaacga cgatatggcg ataggtgcga tgaaagcgtt acacgagcgc
     2821 ggcgtagcgg tgccagagca ggtgtcagtt atcggattcg atgatatcgc tattgccccc
     2881 tacaccgttc cggcgctctc cagcgtaaaa attccggtga ctgagatgat tcaggaaatt
     2941 attggacggc tgatttttat gctcgatggc ggggatttct caccgccgaa aaccttcagc
     3001 ggaaaactga tccgccgcga ctcactcatt gctccttcgc gataataacc tgaggccgat
     3061 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
     3121 gtgacctatc ccattaggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt
     3181 actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt
     3241 ttgatggcgt tggaattccc ggggatccgt cgacctgcag ccaagcttgg cactggccgt
     3301 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3361 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3421 acagttgcgc agcctgaatg gcgaatgccg ttaacggatt cttgaagacg aaagggcctc
     3481 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3541 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3601 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3661 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3721 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3781 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3841 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3901 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3961 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4021 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4081 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4141 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4201 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4261 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4321 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4381 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4441 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4501 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4561 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4621 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4681 aag
//
