LOCUS       sequence-330579-        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             39..1229
                     /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(391..410)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             1868..2059
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2053..2075)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    2991..3010
                     /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     3702..3721
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3802..3819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3973..4561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4053..4072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4732..5523,5524..5592))
                     /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     5355..5374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5593..5697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5765..5783
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5810..5815,5816..5832,5833..5838)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 tatcacagtt aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca
       61 tcgtcatcct cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac
      121 tgccgggcct cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc
      181 tgctagcgct atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg
      241 accgctttgg ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg
      301 cgatcatggc gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca
      361 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
      421 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
      481 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
      541 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
      601 agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
      661 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
      721 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
      781 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
      841 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
      901 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
      961 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     1021 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     1081 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     1141 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     1201 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     1261 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     1321 cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
     1381 gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     1441 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     1501 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     1561 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     1621 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     1681 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     1741 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     1801 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     1861 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     1921 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     1981 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     2041 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2101 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggca ggtgtcgggg
     2161 cgcagccatg acccagtcac cccatggtgc agtatgaagg cggcggagcc gacaccacgg
     2221 ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctt
     2281 tatcaaaaag agtattgact taaagtctaa cctataggat acttacagcg atggagaggt
     2341 gtagtggtaa ccagaagata atatggcttt cgctacctgg agagacgcgc ccgctgatcc
     2401 tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa tactggcagg
     2461 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
     2521 tgattaaata tgatgaaaac ggcaacccgt ggtaccggct tacggcggtg atttttgcga
     2581 tacgccgaac gaaatgtgag cgagtaacaa ccatcgccag ttctgtatga acggtctggt
     2641 ctttgccgac cgcacgccgc atccagcgct gacggaagca aaacaccagc agcagttttt
     2701 cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc
     2761 gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc aagcggtgaa
     2821 gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgtctga actaccgcag
     2881 ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca
     2941 tggtcagaag ccgggcacat cagcgcctgg cagcagtagc gtctggcgga aattgtgagc
     3001 gctcacaatt aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga
     3061 ccaccagcga aatggatttt tgcactcgag ctgggtaata agcgttggca atttaaccgc
     3121 cagtcaggct ttctttcaca gatgtggatt ggcgataaaa aacaactgct gacgccgctg
     3181 cgcgatcagt tcacccgtgc accgctggat aacgacattg gcgtaagtga agcgacccgc
     3241 attgacccta acgcctgggt cgaacactgg aaggcggcgg gccattacca ggccgaagca
     3301 gcgttgttgc agtgcacggc agatacactt gctgatgcgg tgctgattac gaccgctcac
     3361 gcgtggcagc atcaggggaa aaccttattt atcagccgga aaacctaccg gattgatggt
     3421 agtggtcaaa tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg
     3481 cggattggcc tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta
     3541 gcggccgcaa gaaaactatc ccgaccgcct tactgccgcc tgttttgacc gctgggatct
     3601 gctgtaacag agcattagcg caaggtgatt tttgtcttct tgcgctaatt ttttccattg
     3661 tctagagtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     3721 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3781 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4381 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5041 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5341 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5701 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5761 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttctcatgtt tgacagctta
     5821 tcatcgataa gctttaatgc ggtagtt
//
