LOCUS       sequence-436472-        4249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a chimeric LacI:ScrR transcription 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4249)
  AUTHORS   O'Neil P
  TITLE     Expanding the Toolbox of LacI/GalR Family Chimeras for 
            Transcriptional Regulation
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4249)
  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            "unpublished"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: LLhScrR-pHG165a 3mut
FEATURES             Location/Qualifiers
     source          1..4249
                     /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             1597..1788
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1782..1804)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1890..2030
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1945..1964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2045..2062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2216..2804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2296..2315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2975..3766,3767..3835))
                     /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     3598..3617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3836..3940)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4008..4026
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4179..4201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4193..4210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4194..4210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
       61 attcagggtg gtgaatatga aaccggttac tttatacgac gttgccgaat atgccggtgt
      121 gagttatcag accgtgagtc gcgtggtgaa tcaagctagc cacgttagcg ccaaaacccg
      181 tgaaaaagtg gaagccgcca tggccgaact gaactatatc ccgaatcgca ttgcccagca
      241 actggccggt aaacagagct cactgatcgg tttagtgttc ccgagtgtta gccatccgtt
      301 ctttggcgaa ctgatcgaaa ctttagaacg caagctgttc gtgcaaggtt acaaagtgat
      361 tttatgcgac agcgagaaag atccggaaaa agaacgcgcc tatctgcgca tgctggccgc
      421 caacaaagtc gattccgtga tcaccggcag tcacaatctg gccatcaacg agtatgagaa
      481 cgtgtcttta ccgatcgtga gttttgaccg ctttctggca ccgggcattc cgatcgtgag
      541 cagccagaac tttcaaggtg gccagaaggc aaccgaagct ttatttgcca gtggcgccca
      601 gaagatcgcc atcattaccg gtgccaacaa taccggcgcc ccgagtgatt atcgtctggc
      661 tggttacaag cagaccatgg aaaagtacgg cgccgagaag accatcttac agattgacaa
      721 tggcaccagc accactttaa agaatttaga aatcgagcgc ttactgcaga acaagaccgt
      781 ggacggcatt ttctgcaccg atgatctgac cgccatcacc gtgatgaaca ttgcccagaa
      841 attaaaaatc agcattccgg aggagctgaa ggtgattggc tacgacggca ccaaactgat
      901 caaacgcatt gcccctcagc tgagcaccat tgtgcaaccg atcgacgaga tgtgtgacgt
      961 gatgatcgat ctgctgctgc gtcgtatgaa ggacccggat gtggctttag aggaaaacta
     1021 tccgattccg atccagctgt ctttaagcga gagttgttga tgagaattcg taataatata
     1081 gctctagatt aattgcgccc gttaacggcg ggcagcgttg ggtcctggcc acgggtgcgc
     1141 atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag
     1201 cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg
     1261 acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg
     1321 aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc
     1381 tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
     1441 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
     1501 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
     1561 cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc
     1621 cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag
     1681 ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt
     1741 taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1801 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1861 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     1921 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     1981 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     2041 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2101 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2161 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2221 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2281 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2341 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2401 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2461 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2521 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2581 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2641 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2701 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2761 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2821 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2881 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2941 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3001 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3061 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3121 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3181 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3241 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3301 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3361 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3421 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3481 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3541 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3601 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3661 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3721 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3781 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3961 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4021 cgaggccctt tcgtcttcaa gaatccgtta acggcattcg ccattcaggc tgcgcaactg
     4081 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4141 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     4201 gacggccagt gccaagcttg gctgcaggtc gacggatccc cgggaattc
//
