LOCUS       sequence-336813-        5847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To amplify transcription template containing LacI operators .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5847)
  TITLE     Vectors to amplify transcription template containing LacI operators
REFERENCE   2  (bases 1 to 5847)
  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: pRS_2S_400
FEATURES             Location/Qualifiers
     source          1..5847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1191
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(353..372)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             1830..2021
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2015..2037)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    2953..2972
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     3664..3683
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3764..3781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3935..4523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4015..4034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4694..5485,5486..5554))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5317..5336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5555..5659)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5727..5745
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5772..5777,5778..5794,5795..5800)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 atgaaatcta acaatgcgct catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc
       61 ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca ttccgacagc
      121 atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca
      181 cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct cgcttcgcta
      241 cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg gatcctctac
      301 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
      361 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
      421 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
      481 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
      541 atgcaggagt cgcataaggg agagcgtcga ccgatgccct tgagagcctt caacccagtc
      601 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
      661 atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc
      721 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
      781 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
      841 atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc
      901 tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg
      961 caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc
     1021 gcggctctta ccagcctaac ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc
     1081 gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct ataccttgtc
     1141 tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg aatggaagcc
     1201 ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa ttcttgcgga
     1261 gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc gccatctcca
     1321 gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg
     1381 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
     1441 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
     1501 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
     1561 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
     1621 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc
     1681 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat
     1741 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga
     1801 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca
     1861 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
     1921 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
     1981 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2041 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2101 cgggtgttgg caggtgtcgg ggcgcagcca tgacccagtc accccatggt gcagtatgaa
     2161 ggcggcggag ccgacaccac ggccaccgat attatttgcc cgatgtacgc gcgcgtggat
     2221 gaagaccagc ccttcccggc tttatcaaaa agagtattga cttaaagtct aacctatagg
     2281 atacttacag cgatggagag gtgtagtggt aaccagaaga taatatggct ttcgctacct
     2341 ggagagacgc gcccgctgat cctttgcgaa tacgcccacg cgatgggtaa cagtcttggc
     2401 ggtttcgcta aatactggca ggcgtttcgt cagtatcccc gtttacaggg cggcttcgtc
     2461 tgggactggg tggatcagtc gctgattaaa tatgatgaaa acggcaaccc gtggtaccgg
     2521 cttacggcgg tgatttttgc gatacgccga acgaaatgtg agcgagtaac aaccatcgcc
     2581 agttctgtat gaacggtctg gtctttgccg accgcacgcc gcatccagcg ctgacggaag
     2641 caaaacacca gcagcagttt ttcagttccg tttatccggg caaaccatcg aagtgaccag
     2701 cgaatacctg ttccgtcata gcgataacga gctcctgcac tggatggtgg cgctggatgg
     2761 taagccgctg gcaagcggtg aagtgcctct ggatgtcgct ccacaaggta aacagttgat
     2821 tgaactgtct gaactaccgc agccggagag cgccgggcaa ctctggctca cagtacgcgt
     2881 agtgcaaccg aacgcgaccg catggtcaga agccgggcac atcagcgcct ggcagcagta
     2941 gcgtctggcg gaaattgtga gcgctcacaa ttaaacctca gtgtgacgct ccccgccgcg
     3001 tcccacgcca tcccgcatct gaccaccagc gaaatggatt tttgcactcg agctgggtaa
     3061 taagcgttgg caatttaacc gccagtcagg ctttctttca cagatgtgga ttggcgataa
     3121 aaaacaactg ctgacgccgc tgcgcgatca gttcacccgt gcaccgctgg ataacgacat
     3181 tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg gtcgaacact ggaaggcggc
     3241 gggccattac caggccgaag cagcgttgtt gcagtgcacg gcagatacac ttgctgatgc
     3301 ggtgctgatt acgaccgctc acgcgtggca gcatcagggg aaaaccttat ttatcagccg
     3361 gaaaacctac cggattgatg gtagtggtca aatggcgatt accgttgatg ttgaagtggc
     3421 gagcgataca ccgcatccgg cgcggattgg cctgaactgc cagctggcgc aggtagcaga
     3481 gcgggtaaac tggctcggat tagcggccgc aagaaaacta tcccgaccgc cttactgccg
     3541 cctgttttga ccgctgggat ctgctgtaac agagcattag cgcaaggtga tttttgtctt
     3601 cttgcgctaa ttttttccat tgtctagagt agcgatagcg gagtgtatac tggcttaact
     3661 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     3721 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     3781 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3841 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3901 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3961 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4021 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4081 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4141 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4201 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4261 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4321 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4381 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4441 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4501 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4561 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4621 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4681 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4741 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4801 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4861 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4921 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4981 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     5041 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5101 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5161 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5221 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5281 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     5341 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5401 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5461 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5521 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5581 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5641 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5701 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     5761 aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta
     5821 aattgctaac gcagtcaggc accgtgt
//
