LOCUS       sequence-45658-e        7345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7345)
  TITLE     Bishop Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7345)
  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            "Unpublished"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: pNRB65
FEATURES             Location/Qualifiers
     source          1..7345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1622..2029
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2030..3124
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     5037..5053
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5037..5053
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5107..5123)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5108..5124)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5157..5177)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5157..5175)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5196..5212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5196..5212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5209..5231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5220..5236
                     /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)."
     promoter        complement(join(5244..5250,5251..5268,5269..5274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5427..5444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5598..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5678..5697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6357..7148,7149..7217))
                     /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     6980..6999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7218..7322)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaatatgaaa cctataggta cggtggccat tctatgtctg
      721 atcccggtac tacctacaga actagagacg agattcagca tatgagatcc aagaacgatc
      781 caattgctgg tcttaagatg catttgattg atctaggtat tgccactgaa gctgaagtca
      841 aagcttacga caagtccgct agaaaatacg ttgacgaaca agttgaatta gctgatgctg
      901 ctcctcctcc agaagccaaa ttatccatct tgtttgaaga cgtctacgtg aaaggtacag
      961 aaactccaac cctaagaggt aggatccctg aagatacttg ggacttcaaa aagcaaggtt
     1021 ttgcctctag ggattaatta aatcgtaagg aaaaataaaa taatagtgct gtgatcgcat
     1081 gatattcttc cctggaagcg ccattttata gcaagaaatg taagtcaagt atattttaac
     1141 tgtatatcaa caatatagct cttttttatg ccttgttgtt tttcttcggg ttttccccac
     1201 acattgtgtg gagagatagt tattaacaga ccggaaaata gccgcccaag gataaacttt
     1261 tatataaagg gaagggtagt tgacccaaaa aatttggatt ctacttttcc agatttactt
     1321 tcaccctttt atatttgctg tagtctgtta tgccaatcag gaaagcattt gaacaaatag
     1381 gtcgaggaga acttctagta tatctacata cctaatatta ttgccttatt aaaaatggaa
     1441 tcccaacaat tacatcaaaa tccacattct cttcaaaatc aattgtcctg tacttccttg
     1501 ttcatgtgtg ttcaaaaacg ttatatttat aggataatta tactctattt ctcaacaagt
     1561 aattggttgt ttggccgagc ggtctaaggc gcctgattca agaaatatct tgaccgcagt
     1621 taactgtggg aatactcagg tatcgtaaga tgcaagagtt cgaatctctt agcaaccatt
     1681 atttttttcc tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga
     1741 tgactggaaa ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga
     1801 aaaattggga gaaaaaggaa aggtgagagc cgcggaaccg gcttttcata tagaatagag
     1861 aagcgttcat gactaaatgc ttgcatcaca atacttgaag ttgacaatat tatttaagga
     1921 cctattgttt tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt
     1981 ttacatttca gcaatatata tatatatatt tcaaggatat accattctaa tgtctgcccc
     2041 taagaagatc gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat
     2101 taaggttctt aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca
     2161 tttaattggt ggtgctgcta tcgatgctac aggtgtccca cttccagatg aggcgctgga
     2221 agcctccaag aaggttgatg ccgttttgtt aggtgctgtg ggtggtccta aatggggtac
     2281 cggtagtgtt agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc
     2341 caacttaaga ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc
     2401 acaatttgct aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt
     2461 tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt
     2521 tccagaagtg caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc
     2581 attgcctatt tggtccttgg ataaagctaa tgttttggcc tcttcaagat tatggagaaa
     2641 aactgtggag gaaaccatca agaacgaatt ccctacattg aaggttcaac atcaattgat
     2701 tgattctgcc gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac
     2761 cagcaacatg tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg
     2821 tttgttgcca tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta
     2881 cgaaccatgc cacggttctg ctccagattt gccaaagaat aaggtcaacc ctatcgccac
     2941 tatcttgtct gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc
     3001 cattgaagat gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg
     3061 ttccaacagt accacggaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc
     3121 ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg aaattcataa
     3181 tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagacgaa actatatacg
     3241 caatctacat acatttatca agaaggagaa aaaggaggat gtaaaggaat acaggtaagc
     3301 aaattgatac taatggctca acgtgataag gaaaaagaat tgcactttaa cattaatatt
     3361 gacaaggagg agggcaccac acaaaaagtt aggtgtaaca gaaaatcatg aaactatgat
     3421 tcctaattta tatattggag gattttctct aaaaaaaaaa aaatacaaca aataaaaaac
     3481 actcaatgac ctgaccattt gatggagttt aagtcaatac cttcttgaac catttcccat
     3541 aatggtgaaa gttccctcaa gaattttact ctgtcagaaa cggccttaac gacgtagtcg
     3601 accaattggt gaaaaaggta ttaccgattc aagtgactag tttttgtata cttttttaat
     3661 gaagatgaca ttgctccttt attaaacttt tcttttactt tgtgttacta atattattaa
     3721 tatcttgtta tgattctttg ttttagcagc tccaaaagaa aggaatatta taatataatt
     3781 gttgtaattc ggaaaaaaaa tattaaatta ctatggtgca tatgcgtata taaaattgag
     3841 cattacctaa cgccatatat tactatctaa agtcctcaca gatgatgtgt tttcaaaatc
     3901 aaatgattgg agctcctcat catcatcccc ggtaaactgc tcggtttcag atccattatc
     3961 cagatacctc aacgggctag tgtacgcaat tgaagggtac ctataaaata aagtacgttc
     4021 ctctttcaac tgaaaaaacc cccgataatc ccttgacaag aaataataca taatataatc
     4081 tcgctttctg acaagacttt tcagttcttg gtactctaat tcccgttcga aaaccggata
     4141 tgtgccagca atttcgtcag ttattatagc gtttataata gcctcgtagt catcgggctc
     4201 aattctattt tcgtagttga acagacggta aaaagggcta cagtcataat tatgctcgca
     4261 ttcctcgaga ataatatctg aatcagattg taaatgtatg atatttcttg gcgttacaac
     4321 aatagaatcc tcaggccctg atattttctc tatagatggg gaatccggta tatagtcaag
     4381 gttgatatct gttaattcat ttgaatttag gttagcgcga gcttcaattt cgttaggctt
     4441 cttgaaattt gccttaactg ggatttgctt tgccaatttt ttgtcatcta gcttttttct
     4501 atgattgaaa tttaggtcac gaagccttat atgttcaaga tccttggatt ttgtagcagc
     4561 tttttccttt atcaagaaac tgcttgactt cttttgcgaa aaagggtggg tttgattttc
     4621 atcttgatgc aacctctcat cgacatccat tttgtcactg tgtctgagaa tctatacttc
     4681 agggcctgct acaagaagta gaaactaaaa tacaatgtta caaatgcgag aagggttttt
     4741 tttatgatca tgagtgaccg caataaaatc tttcgaagct ttttcgaata ttttaacatt
     4801 ttttctctat agcttctttg tggcatcgcc catgggatca aaccatactg gtttctttgt
     4861 aaaacaggcg caaaatatag aaactcagaa aaagagaaaa cgagtagtgg aaaaatttat
     4921 cagccattta tgcgacgcat ccgtttgact gatctaccta tttaaaacag aaacctacgt
     4981 tgaaaagtta tttttccaac acattactgg atctcatcaa attgccaatg taatcttcta
     5041 gaactagtgg atccggtccg gttaccttgg gggggggggg ctgcaggaat tcgatatcaa
     5101 gcttatcgat accgtcgacc tcgaggccta tagggcccgg tacccagctt ttgttccctt
     5161 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5221 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5281 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5401 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6001 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7321 cgcacatttc cccgaaaagt gccac
//
