LOCUS       sequence-336805-        5848 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 5848)
  TITLE     Vectors to amplify transcription template containing LacI operators
REFERENCE   2  (bases 1 to 5848)
  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: pYY_N1_400
FEATURES             Location/Qualifiers
     source          1..5848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    84..100
                     /label=lac operator
                     /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)."
     primer_bind     794..813
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(894..911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1065..1653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1145..1164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1824..2615,2616..2684))
                     /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     2447..2466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2685..2789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2857..2875
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2902..2907,2908..2924,2925..2930)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2978..4168
                     /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(3330..3349)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             4807..4998
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4992..5014)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agtacgcgta gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca tcagcgcctg
       61 gcagcagtag cgtctggcgg aaattgtgag cggataacaa ttaaacctca gtgtgacgct
      121 ccccgccgcg tcccacgcca tcccgcatct gaccaccagc gaaatggatt tttgcactcg
      181 agctgggtaa taagcgttgg caatttaacc gccagtcagg ctttctttca cagatgtgga
      241 ttggcgataa aaaacaactg ctgacgccgc tgcgcgatca gttcacccgt gcaccgctgg
      301 ataacgacat tggcgtaagt gaagcgaccc gcattgaccc taacgcctgg gtcgaacact
      361 ggaaggcggc gggccattac caggccgaag cagcgttgtt gcagtgcacg gcagatacac
      421 ttgctgatgc ggtgctgatt acgaccgctc acgcgtggca gcatcagggg aaaaccttat
      481 ttatcagccg gaaaacctac cggattgatg gtagtggtca aatggcgatt accgttgatg
      541 ttgaagtggc gagcgataca ccgcatccgg cgcggattgg cctgaactgc cagctggcgc
      601 aggtagcaga gcgggtaaac tggctcggat tagcggccgc aagaaaacta tcccgaccgc
      661 cttactgccg cctgttttga ccgctgggat ctgctgtaac agagcattag cgcaaggtga
      721 tttttgtctt cttgcgctaa ttttttccat tgtctagagt agcgatagcg gagtgtatac
      781 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
      841 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
      901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      961 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1021 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1081 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1141 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1201 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1261 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1321 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1381 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1441 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1501 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1561 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1621 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1681 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1741 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1801 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1861 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1921 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1981 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2041 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2101 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     2161 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2221 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2281 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2341 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2401 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     2461 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2521 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2581 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2641 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2701 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2761 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     2821 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     2881 cgtcttcaag aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt
     2941 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
     3001 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
     3061 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct
     3121 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga
     3181 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
     3241 gatcatggcg accacacccg tcctgtggat cctctacgcc ggacgcatcg tggccggcat
     3301 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     3361 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     3421 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
     3481 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     3541 gcgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3601 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3661 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3721 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     3781 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     3841 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     3901 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     3961 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     4021 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     4081 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     4141 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     4201 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     4261 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
     4321 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     4381 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     4441 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     4501 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     4561 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     4621 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     4681 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     4741 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     4801 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     4861 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     4921 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
     4981 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5041 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcag gtgtcggggc
     5101 gcagccatga cccagtcacc ccatggtgca gtatgaaggc ggcggagccg acaccacggc
     5161 caccgatatt atttgcccga tgtacgcgcg cgtggatgaa gaccagccct tcccggcttt
     5221 atcaaaaaga gtattgactt aaagtctaac ctataggata cttacagcga tggagaggtg
     5281 tagtggtaac cagaagataa tatggctttc gctacctgga gagacgcgcc cgctgatcct
     5341 ttgcgaatac gcccacgcga tgggtaacag tcttggcggt ttcgctaaat actggcaggc
     5401 gtttcgtcag tatccccgtt tacagggcgg cttcgtctgg gactgggtgg atcagtcgct
     5461 gattaaatat gatgaaaacg gcaacccgtg gtaccggctt acggcggtga tttttgcgat
     5521 acgccgaacg tcgatcgagc tcaacgcaat tatcgccagt tctgtatgaa cggtctggtc
     5581 tttgccgacc gcacgccgca tccagcgctg acggaagcaa aacaccagca gcagtttttc
     5641 agttccgttt atccgggcaa accatcgaag tgaccagcga atacctgttc cgtcatagcg
     5701 ataacgagct cctgcactgg atggtggcgc tggatggtaa gccgctggca agcggtgaag
     5761 tgcctctgga tgtcgctcca caaggtaaac agttgattga actgtctgaa ctaccgcagc
     5821 cggagagcgc cgggcaactc tggctcac
//
