LOCUS       sequence-178295-        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 Q55A
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..94
                     /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(114..133)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1358..1375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1358..1374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1367..1389)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1542..1560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1628..1732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1733..1801,1802..2593)
                     /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(1951..1970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2764..3352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3253..3272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3506..3523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3538..3678
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3604..3623)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3764..3786
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3780..3971)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4571..4592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4607..4612,4613..4630,4631..4637)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     4667..4683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4667..4683
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata
       61 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
      121 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
      181 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctca attacattcc
      241 caaccgcgtg gcacaagccc tggcgggcaa acagagccag acgctggggc tggtagtgac
      301 caacacgctt taccacggca tttattttag tgaattactc tttcatgccg cgcgaatggc
      361 ggaagagaaa gggcggcagt tgctattggc agatggtaaa cacagcgcag aagaagagcg
      421 ccaggcgatt cagtatctgc tggatctgcg ctgcgacgcg atcatgattt acccgcgttt
      481 tttaagcgtg gatgagatcg atgacatcat tgacgcgcac agtcagccga taatggtgct
      541 taatcgccgc ctgcgcaaaa acagcagcca tagcgtctgg tgcgatcata aacagaccag
      601 ttttaacgcc gtggcagagt tgataaacgc cgggcatcag gagattgctt tccttaccgg
      661 ctcgatggat tcccccacca gcattgaacg tcttgccggg tataaagacg cgctggcgca
      721 gcatggtatt gcgctcaatg aaaaacttat cgctaacggt aaatggacgc ctgccagcgg
      781 ggccgaaggg gtagaaatgt tgctcgaacg tggggctaaa tttagcgcgt tagttgccag
      841 taacgacgat atggcgatag gtgcgatgaa agcgttacac gagcgcggcg tagcggtgcc
      901 agagcaggtg tcagttatcg gattcgatga tatcgctatt gccccctaca ccgttccggc
      961 gctctccagc gtaaaaattc cggtgactga gatgattcag gaaattattg gacggctgat
     1021 ttttatgctc gatggcgggg atttctcacc gccgaaaacc ttcagcggaa aactgatccg
     1081 ccgcgactca ctcattgctc cttcgcgata ataacctgag gccgatactg tcgtcgtccc
     1141 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat
     1201 taggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     1261 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga
     1321 attcccgggg atccgtcgac ctgcagccaa gcttggcact ggccgtcgtt ttacaacgtc
     1381 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1441 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1501 tgaatggcga atgccgttaa cggattcttg aagacgaaag ggcctcgtga tacgcctatt
     1561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2881 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3541 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     3601 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3661 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3721 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3781 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3841 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3901 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     3961 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4021 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4081 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     4141 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     4201 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     4261 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     4321 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     4381 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     4441 caggacccaa cgctgcccgc cgttaacggc caatacgcaa accgcctctc cccgcgcgtt
     4501 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     4561 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     4621 ttccggctcg tatgttgtgt ggcgtggtac tcataacaat ttcacacagg aaacagctat
     4681 gac
//
