LOCUS       sequence-436470-        4306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:SalR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4306)
  AUTHORS   O'Neil P
  TITLE     Expanding the Toolbox of LacI/GalR Family Chimeras for 
            Transcriptional Regulation
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4306)
  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"; journalName: 
            "unpublished"
COMMENT     Sequence Label: LLhSalR-pHG165a 3mut
FEATURES             Location/Qualifiers
     source          1..4306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      194..203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1654..1845
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1839..1861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1947..2087
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2002..2021
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2102..2119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2273..2861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2353..2372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3032..3823,3824..3892))
                     /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     3655..3674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3893..3997)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4065..4083
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4236..4258
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4250..4267
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4251..4267
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
       61 attcagggtg gtgaatatga aacccgttac tttatatgat gtggccgagt atgctggtgt
      121 gagctatcag accgtgagcc gtgttgtgaa ccaagctagc catgtgagcg ccaaaacccg
      181 cgagaaagtt gaagccgcca tggccgaact gaactacatt ccgaatcgcc tcgcccaggg
      241 tctggctggt aagcagagca ggatgatggc ctttgtgctg ggcagcaatt tagataatcc
      301 gagcgaatct ttacagaacg aatacattag cgctttacaa gttggtgcag tgatggcagc
      361 ccgtgaagcc ggttatcatt tactggtgga accgctgggt atggcagccg aaggttttga
      421 agaacgcacc cgtcgtctga ttgcaatgcc ggcagtggac ggctttattt tcatgccggg
      481 tttagcagat aacccggtgc tgctggaact gctggttcag cgcccgggta cctatgtgcg
      541 cgttagcccg ggtcgtgaag tgccgagctt acccgctatg gtgcgcatcg acgactatca
      601 agctgccttc gatatgaccc gccatctgct ggatctgggt catcgtcgca ttgccttcat
      661 tcaaggtctg ccggattttg gtagcgccga agcccgcttt aacgcctttt gcgccgcaat
      721 ggcaccgcat ggtggtcatg atccggcact gctgcgcagc ggtgattatc gtctggccag
      781 cggtcgcgaa gccgcagaaa gcttactgag ccatcccgaa ccgccgacag caatctttgc
      841 cgcaaatgat gccatggcac tgggtgcaat ggcagttgcc catcaccgcg gcattcgcgt
      901 gccggatgat ctgagtatcg ccggtatcga cgatatcccg atggccgccg cagtttggcc
      961 gagtctgacc accgttcgtc agccgctggt tgccatgggt aaagccgccg cccactacat
     1021 tatcgcccgt agtgagggtc gccatgatgg tccgctgccc ggtacctttc gccatgaagt
     1081 ggtgatgcgt gaaagcaccg ccccgccgcg cggctgatga gaattcgtaa taatatagct
     1141 ctagattaat tgcgcccgtt aacggcgggc agcgttgggt cctggccacg ggtgcgcatg
     1201 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     1261 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     1321 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     1381 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     1441 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     1501 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     1561 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     1621 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     1681 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     1741 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     1801 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1861 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1921 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     1981 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     2041 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     2101 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2161 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2221 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2281 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2341 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2401 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2461 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2521 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2581 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2641 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2701 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2761 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2821 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2881 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2941 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3001 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3061 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3121 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3181 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3241 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3301 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3361 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3421 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3481 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3541 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3601 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3661 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3721 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3781 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3841 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3901 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3961 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4021 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4081 ggccctttcg tcttcaagaa tccgttaacg gcattcgcca ttcaggctgc gcaactgttg
     4141 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     4201 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     4261 ggccagtgcc aagcttggct gcaggtcgac ggatccccgg gaattc
//
