LOCUS       sequence-50976-e        6376 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 6376)
  AUTHORS   Chee MK, Haase SB
  TITLE     New and Redesigned pRS Plasmid Shuttle Vectors for Genetic 
            Manipulation of Saccharomycescerevisiae.
  JOURNAL   G3 (Bethesda). 2012 May;2(5):515-26. Epub 2012 May 1.
  PUBMED    22670222
REFERENCE   2  (bases 1 to 6376)
  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: "G3 
            (Bethesda)."; date: "2012-05"; volume: "2(5)"; pages: "515-26. Epub 
            2012 May 1"
COMMENT     Sequence Label: pRSII429
FEATURES             Location/Qualifiers
     source          1..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2002..2457)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2139..2160)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2351..2370
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2434..2814)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2583..2605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2597..2614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2598..2614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2624..2643
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2624..2642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2651..2758
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2667..2683
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2668..2684
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2718..2734)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2718..2734)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2771..2791)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2771..2789)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2810..2826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2810..2826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2823..2845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2834..2850
                     /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(2858..2864,2865..2882,2883..2888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2903..2924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3041..3058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3212..3800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3292..3311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3971..4762,4763..4831))
                     /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     4594..4613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4832..4936)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4963..6309
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6349..6367
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgcga gtcggcgagc aaacagggtg ctgctactaa
      361 acgaacaata aagaaagaca atttgggcac ccaatataaa atgagcagtc aattcaatca
      421 attcagtgtg aaattttttt cttggaaaga tcattcactt ctacacagct gtacatgagt
      481 ttatatgaca gttttttttt atgtacatcc agttcaagat aatcagatta tatattggcc
      541 caaaagggat cattaacgaa ttgcgaaatg ggtaatctct ttacagttaa cagattctct
      601 ggtgattggc tttcctcaag gtagcaaata tactctaatt gtagcacgtc tactatgtat
      661 ttctttacct tgtcatagca cacacccact tgcgccacgc cgtgtgcgtc atcacttaga
      721 acaaatctgg agccacagtg cttcttgacc agattacata aggttttgct ggggtacggc
      781 tcctccaggc gctttcttaa tgcggacgta ttgatttcaa ttgcgccgcc ataggagtct
      841 ataaattgta aatttcttac aactgcatcg tatatttctg gccattcact gatgacgtcc
      901 agtgaagcta caggaactcc ggtttcttcg ttgcagttgc ccgatttctg gtttactagc
      961 atgtcattgg gcaaaaataa tttgtaaagg tcgaagtgac ccacgaccaa cggtttaata
     1021 ttgatcagca tttcgtactg tgattggaag taagacagga gaaaatgttt caaattatca
     1081 ttgaaggaat gcaatgaatt gtaccattgt tgttggtcga aatcaatagg tatcccgttg
     1141 acgtgatgga ccgaacccac acaaaacttc aaaatatcat tattctcctt catgagtcgc
     1201 tttgcatatt cgatatgagc catgtcacaa ctttcgatct ccattcctat aatgaattta
     1261 gtccgcacat cgggtctatc agcataacga gtcttgattt cttgcgcatg actcatgaaa
     1321 ttcttgaacg atgtttctag cttggttatg acttcctcag gattcttgcc caatgactgc
     1381 tcttcggggt atataaactt ggcctcaatt cttggtatgt gctctgtcaa acagtacgtg
     1441 tgaaagttga ggttgaccac ttgatcgacc acggaatcca aagggtccgt accgtgggca
     1501 ctatagtcac cggagtgtga atggtgtgag tgcatgacct tgaggatttt tactgaattg
     1561 tacgtatcaa gaagtactaa aatgtgtttg aaagggaaag attgtacggg aaatggaaaa
     1621 tgtacatata atatccagta tcatttgagt tgatagagat tatctctata gatagagaga
     1681 tttgacaaaa aaaataaaaa aatttaaatg gacgagtccg gaatcgaacc ggagacctct
     1741 cccatgctaa gggagcgcgc taccgactac gccacacgcc cgaacgtttg ttgaaatctt
     1801 ccaaacagaa atgatacagg tttactttac agcaagacat tagtagtttc attagcttat
     1861 caacatgtat gaaaccggca agattgaaag atgcatgtat actaaactca caaattagag
     1921 cttcaattta attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta
     1981 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2041 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2101 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2161 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2221 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2281 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2341 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2401 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     2461 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2521 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     2581 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     2641 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     2701 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     2761 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2821 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2881 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2941 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4861 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4921 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt
     4981 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     5041 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     5101 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     5161 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     5221 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     5281 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     5341 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     5401 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     5461 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     5521 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     5581 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     5641 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     5701 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     5761 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     5821 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     5881 ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     5941 tctagctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc
     6001 ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc
     6061 tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta
     6121 aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga
     6181 tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc
     6241 tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga
     6301 tattggatca tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6361 acgaggccct ttcgtc
//
