LOCUS       sequence-178381-        4713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:GalS transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4713)
  TITLE     Swint-Kruse lab plasmids
REFERENCE   2  (bases 1 to 4713)
  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"
COMMENT     Sequence Label: LShS-pHG165a E230K
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..79
                     /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(99..118)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1373..1390)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1373..1389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1382..1404)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1557..1575)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1643..1747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1748..1816,1817..2608)
                     /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(1966..1985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2779..3367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3268..3287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3521..3538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3553..3693
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3619..3638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3779..3801
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3795..3986)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4586..4607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4622..4627,4628..4645,4646..4652)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     4682..4698
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4682..4698
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
       61 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      121 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      181 gcgggaaaaa gtggaagcgg cgatggcgga gctcgattat cggccaaacg ccaatgccca
      241 ggcgctggca actcaggtta gcgacaccat tggcgtggtg gtgatggacg tttctgatgc
      301 gtttttcggc gcgctggtaa aagcggtgga tctggtcgct cagcagcatc agaaatacgt
      361 gctaatcggc aatagctatc atgaagcgga aaaagagcgt cacgccattg aggtgttaat
      421 tcgccagcgt tgtaatgcgt tgattgttca ctcaaaagca ttgagtgacg atgaactggc
      481 gcaatttatg gataacattc ccggtatggt gttaatcaac cgcgttgtgc cggggtacgc
      541 ccatcgttgc gtttgcctgg ataatctcag cggtgcccga atggcgacgc gcatgttgct
      601 gaataacggt catcaacgta ttggttatct ttcttccagc cacggcattg aagatgacgc
      661 catgcgtaaa gcaggctgga tgagtgcgtt gaaagagcag gatattattc cgccggaaag
      721 ctggattggc gctggtacgc cggacatgcc gggcggtgag gcggcgatgg ttaaactgct
      781 ggggcgcaat ctacaactta ccgctgtatt tgcttataac gacaatatgg ccgctggcgc
      841 actgacagca ttaaaagata atggcattgc gattccgtta catctctcaa tcatcggttt
      901 cgatgatatt cccatcgccc gttacaccga cccgcaatta acgaccgtgc gttatcccat
      961 tgcttcaatg gcgaaattag ccaccgaact ggccttgcag ggggcagcag gcaatattga
     1021 tcctcgtgcc agccactgtt ttatgccgac gttagtgcgt cgccattctg tcgcaacacg
     1081 ccagaatgcg gcggcgatca ctaactcaac aaatcaggcg atgtaataac ctcaggccga
     1141 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     1201 cgtgacctat cccattaggt caatccgccg tttgttccca cggagaatcc gacgggttgt
     1261 tactcgctca catttaatgt tgatgaaagc tggctacggg aaggccagac gcgaattatt
     1321 tttgatggcg ttggaattcc cggggatccg tcgacctgca gccaagcttg gcactggccg
     1381 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     1441 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1501 aacagttgcg cagcctgaat ggcgaatgcc gttaacggat tcttgaagac gaaagggcct
     1561 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1621 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1681 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1741 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1801 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1861 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1921 tcgccccgaa gaacgttttc cgatgatgag cacttttaaa gttctgctat gtggcgcggt
     1981 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2041 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2101 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2161 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2221 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2281 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2341 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2401 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2461 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2521 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2581 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2701 tctcatgacc aaaatccctt aacgtaagtt ttcgttccac tgagcgtcag accccgtaga
     2761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2881 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgt tcgtcagggg ggcggagcct
     3361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3541 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3601 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3661 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3721 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3781 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     3841 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     3901 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     3961 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     4021 agggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     4081 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     4141 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aagtgttcca
     4201 cagggtagcc agcagcatcc tgcgatgcag atctggaaca taatggtgca gggcgctgac
     4261 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4321 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4381 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     4441 atgcgcaccc gtggccagga cccaacgctg cccgccgtta acggccaata cgcaaaccgc
     4501 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4561 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4621 ctttacactt tatgcttccg gctcgtatgt tgtgtggcgt ggtactcata acaatttcac
     4681 acaggaaaca gctatgacca tgattacgaa ttc
//
