LOCUS       sequence-178388-        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)
  TITLE     Swint-Kruse lab plasmids
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"
COMMENT     Sequence Label: LAhA-pHG165a
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..81
                     /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(101..120)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(1345..1362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1345..1361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1354..1376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1529..1547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1615..1719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1720..1788,1789..2580)
                     /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(1938..1957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2751..3339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3240..3259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3493..3510
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3525..3665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3591..3610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3751..3773
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3767..3958)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    4558..4579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4594..4599,4600..4617,4618..4624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     4654..4670
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4654..4670
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tccgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag
       61 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
      121 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
      181 acgcgggaaa aagtggaagc ggcgatggcg gagagcggtt accgtccaaa cttgctggcg
      241 cgcaatctgt cggcgaagag tactcagacg ctggggctgg tagtgaccaa cacgctttac
      301 cacggcattt attttagtga attactcttt catgccgcgc gaatggcgga agagaaaggg
      361 cggcagttgc tattggcaga tggtaaacac agcgcagaag aagagcgcca ggcgattcag
      421 tatctgctgg atctgcgctg cgacgcgatc atgatttacc cgcgtttttt aagcgtggat
      481 gagatcgatg acatcattga cgcgcacagt cagccgataa tggtgcttaa tcgccgcctg
      541 cgcaaaaaca gcagccatag cgtctggtgc gatcataaac agaccagttt taacgccgtg
      601 gcagagttga taaacgccgg gcatcaggag attgctttcc ttaccggctc gatggattcc
      661 cccaccagca ttgaacgtct tgccgggtat aaagacgcgc tggcgcagca tggtattgcg
      721 ctcaatgaaa aacttatcgc taacggtaaa tggacgcctg ccagcggggc cgaaggggta
      781 gaaatgttgc tcgaacgtgg ggctaaattt agcgcgttag ttgccagtaa cgacgatatg
      841 gcgataggtg cgatgaaagc gttacacgag cgcggcgtag cggtgccaga gcaggtgtca
      901 gttatcggat tcgatgatat cgctattgcc ccctacaccg ttccggcgct ctccagcgta
      961 aaaattccgg tgactgagat gattcaggaa attattggac ggctgatttt tatgctcgat
     1021 ggcggggatt tctcaccgcc gaaaaccttc agcggaaaac tgatccgccg cgactcactc
     1081 attgctcctt cgcgataata acctgaggcc gatactgtcg tcgtcccctc aaactggcag
     1141 atgcacggtt acgatgcgcc catctacacc aacgtgacct atcccattag gtcaatccgc
     1201 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1261 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt cccggggatc
     1321 cgtcgacctg cagccaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1381 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1441 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1501 ccgttaacgg attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1561 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1741 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1801 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1861 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1921 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1981 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2041 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2101 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2161 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2221 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2281 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2341 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2401 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2461 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2521 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2581 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2881 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3421 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3481 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3541 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3601 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3661 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3721 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3781 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3841 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3901 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3961 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4021 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4081 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4141 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4201 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4261 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4321 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4381 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     4441 tgcccgccgt taacggccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4501 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4561 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4621 gttgtgtggc gtggtactca taacaatttc acacaggaaa cagctatgac catgattacg
     4681 aat
//
