LOCUS       sequence-339114-        5879 bp    DNA     circular SYN 15-APR-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 5879)
  TITLE     Vectors to amplify transcription template containing LacI operators
REFERENCE   2  (bases 1 to 5879)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pYY_S2_400
FEATURES             Location/Qualifiers
     source          1..5879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(569..1360,1361..1429))
                     /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     1192..1211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1430..1534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1602..1620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1647..1652,1653..1669,1670..1675)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1723..2913
                     /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(2075..2094)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3552..3743
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3737..3759)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    4308..4327
                     /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     5418..5437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5518..5535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5689..5879,1..398))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5769..5788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
       61 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      241 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
      901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     1201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1561 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1621 cctttcgtct tcaagaattc tcatgtttga cagcttatca tcgataagct ttaatgcggt
     1681 agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg
     1741 ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg
     1801 gtactgccgg gcctcttgcg ggatatcgtc cattccgaca gcatcgccag tcactatggc
     1861 gtgctgctag cgctatatgc gttgatgcaa tttctatgcg cacccgttct cggagcactg
     1921 tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc tacttggagc cactatcgac
     1981 tacgcgatca tggcgaccac acccgtcctg tggatcctct acgccggacg catcgtggcc
     2041 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     2101 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     2161 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     2221 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     2281 ggagagcgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     2341 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     2401 caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
     2461 atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
     2521 actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc
     2581 gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt
     2641 atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg
     2701 caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta
     2761 acttcgatca ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg
     2821 aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt
     2881 cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg
     2941 gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa
     3001 ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca
     3061 tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga
     3121 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     3181 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     3241 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     3301 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     3361 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     3421 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     3481 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     3541 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     3601 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     3661 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     3721 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3781 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcaggtgtc
     3841 ggggcgcagc catgacccag tcaccccatg gtgcagtatg aaggcggcgg agccgacacc
     3901 atggggtgac tgggtcatgg ctacgccccg acaccacggc caccgatatt atttgcccga
     3961 tgtacgcgcg cgtggatgaa gaccagccct tcccggcttt atcaaaaaga gtattgactt
     4021 aaagtctaac ctataggata cttacagcga tggagaggtg tggtaaccag aagataagat
     4081 ggctttcgct acctggagag acgcgcccgc tgatcctttg cgaatacgcc cacgcgatgg
     4141 gtaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac
     4201 agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca
     4261 acccgtggta ccggcttacg gcggtgattt tggcgatacg ccgaacgaat tgtgagcgct
     4321 cacaattatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac gccgcatcca
     4381 gcgctgacgg aagcaaaaca ccagcagcag tttttccagt tccgtttatc cgggcaaacc
     4441 atcgaagtga ccagcgaata cctgttccgt catagcgata acgagctcct gcactggatg
     4501 gtggcgctgg atggtaagcc gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa
     4561 ggtaaacagt tgattgaact gcctgaacta ccgcagccgg agagcgccgg gcaactctgg
     4621 ctcacagtac gcgtagtgca accgaacgcg accgcatggt cagaagccgg gcacatcagc
     4681 gcctggcagc agtggcgtct ggcggaaaat tgtagcgagt aacaaccaaa cctcagtgtg
     4741 acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat ggattttgca
     4801 ctcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct ttcacagatg
     4861 tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac ccgtgcaccg
     4921 ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc ctgggtcgaa
     4981 cgctggaagg cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat
     5041 acacttgctg atgcggtgct gattacgacc gctcacgcgt ggcagcatca ggggaaaacc
     5101 ttatttatca gccggaaaac ctaccggatt gatggtagtg gtcaaatggc gattaccgtt
     5161 gatgttgaag tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg
     5221 gcgcaggtag cagagcgggt aaactggctc ggattagcgg ccgcaagaaa actatcctga
     5281 ccgccttact gccgcctgtt ttgaccgctg ggatctgctg taacagagca ttagcgcaag
     5341 gtgatttttg tcttcttgcg ctaatttttt ccattgtcta gagtagcgat agcggagtgt
     5401 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5461 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     5521 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5581 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5641 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5701 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5761 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5821 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgcttt
//
