LOCUS       sequence-49273-e        6865 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 6865)
  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 6865)
  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: pRSII422
FEATURES             Location/Qualifiers
     source          1..6865
                     /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"
     promoter        199..574
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     rep_origin      complement(2495..2950)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2632..2653)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2844..2863
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2927..3307)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3076..3098
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3090..3107
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3091..3107
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3117..3136
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3117..3135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3144..3251
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3160..3176
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3161..3177
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3211..3227)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3211..3227)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3264..3284)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3264..3282)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3303..3319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3303..3319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3316..3338)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3327..3343
                     /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(3351..3357,3358..3375,3376..3381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3396..3417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3534..3551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3705..4293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3785..3804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4464..5255,5256..5324))
                     /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     5087..5106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5325..5429)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5456..6802
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6842..6860
                     /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 accatagatc tgaattaatt cttgaataat acataacttt tcttaaaaga atcaaagaca
      241 gataaaattt aagagatatt aaatattagt gagaagccga gaattttgta acaccaacat
      301 aacactgaca tctttaacaa cttttaatta tgatacattt cttacgtcat gattgattat
      361 tacagctatg ctgacaaatg actcttgttg catggctacg aaccgggtaa tactaagtga
      421 ttgactcttg ctgacctttt attaagaact aaatggacaa tattatggag catttcatgt
      481 ataaattggt gcgtaaaatc gttggatctc tcttctaagt acatcctact ataacaatca
      541 agaaaaacaa gaaaatcgga caaaacaatc aagtatggat tcaagaacag ttggtatatt
      601 aggaggggga caattgggac gtatgattgt tgaggcagca aacaggctca acattaagac
      661 ggtaatacta gatgctgaaa attctcctgc caaacaaata agcaactcca atgaccacgt
      721 taatggctcc ttttccaatc ctcttgatat cgaaaaacta gctgaaaaat gtgatgtgct
      781 aacgattgag attgagcatg ttgatgttcc tacactaaag aatcttcaag taaaacatcc
      841 caaattaaaa atttaccctt ctccagaaac aatcagattg atacaagaca aatatattca
      901 aaaagagcat ttaatcaaaa atggtatagc agttacccaa agtgttcctg tggaacaagc
      961 cagtgagacg tccctattga atgttggaag agatttgggt tttccattcg tcttgaagtc
     1021 gaggactttg gcatacgatg gaagaggtaa cttcgttgta aagaataagg aaatgattcc
     1081 ggaagcattg gaagtactga aggatcgtcc tttgtacgcc gaaaaatggg caccatttac
     1141 taaagaatta gcagtcatga ttgtgaggtc tgttaacggt ttagtgtttt cttacccaat
     1201 tgtagagact atccacaagg acaatatttg tgacttatgt tatgcgcctg ctagagttcc
     1261 ggactccgtt caacttaagg cgaagttgtt ggcagaaaat gcaatcaaat cttttcccgg
     1321 ttgtggtata tttggtgtgg aaatgttcta tttagaaaca ggggaattgc ttattaacga
     1381 aattgcccca aggcctcaca actctggaca ttataccatt gatgcttgcg tcacttctca
     1441 atttgaagct catttgagat caatattgga tttgccaatg ccaaagaatt tcacatcttt
     1501 ctccaccatt acaacgaacg ccattatgct aaatgttctt ggagacaaac atacaaaaga
     1561 taaagagcta gaaacttgcg aaagagcatt ggcgactcca ggttcctcag tgtacttata
     1621 tggaaaagag tcaagaccta acagaaaagt aggtcacata aatattattg cctccagtat
     1681 ggcggaatgt gaacaaaggc tgaactacat tacaggtaga actgatattc caatcaaaat
     1741 ctctgtcgct caaaagttgg acttggaagc aatggtcaaa ccattggttg gaatcatcat
     1801 gggatcagac tctgacttgc cggtaatgtc tgccgcatgt gcggttttaa aagattttgg
     1861 cgttccattt gaagtgacaa tagtctctgc tcatagaact ccacatagga tgtcagcata
     1921 tgctatttcc gcaagcaagc gtggaattaa aacaattatc gctggagctg gtggggctgc
     1981 tcacttgcca ggtatggtgg ctgcaatgac accacttcct gtcatcggtg tgcccgtaaa
     2041 aggttcttgt cttgatggag tagattcttt acattcaatt gtgcaaatgc ctagaggtgt
     2101 tccagtagct accgtcgcta ttaataatag tacgaacgct gcgctgttgg ctgtcagact
     2161 gcttggcgct tatgattcaa gttatacaac gaaaatggaa cagtttttat taaagcaaga
     2221 agaagaagtt cttgtcaaag cacaaaagtt agaaactgtc ggttacgaag catatcttga
     2281 aaacaagtaa tatataagtt tattgatata cttgtacagc aaataattat aaaatgatat
     2341 acctattttt taggctttgt tatgattaca tcaaatgtgg acttcataca tagaaatcaa
     2401 cgcttacagg tgtccttttt taagaatttc atacataaga tctatgcggt gtgaaatacc
     2461 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     2521 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2581 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2641 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2701 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2761 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2821 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2881 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2941 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3001 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3061 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     3121 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata
     3181 agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg gccgccaccg
     3241 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     3301 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3361 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3421 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3481 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3541 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3601 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3661 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3721 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3781 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3841 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3901 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3961 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4021 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4081 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4141 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4201 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4261 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4321 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4381 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4441 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4501 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4561 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4621 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4681 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4741 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4801 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4861 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4921 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4981 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5041 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5101 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5161 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5221 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5281 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5341 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5401 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaacga
     5461 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     5521 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     5581 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     5641 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     5701 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     5761 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     5821 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     5881 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     5941 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     6001 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     6061 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     6121 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     6181 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     6241 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     6301 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     6361 cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga
     6421 agttcctata ctttctagct agagaatagg aacttcggaa taggaacttc aaagcgtttc
     6481 cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca catacagctc actgttcacg
     6541 tcgcacctat atctgcgtgt tgcctgtata tatatataca tgagaagaac ggcatagtgc
     6601 gtgtttatgc ttaaatgcgt acttatatgc gtctatttat gtaggatgaa aggtagtcta
     6661 gtacctcctg tgatattatc ccattccatg cggggtatcg tatgcttcct tcagcactac
     6721 cctttagctg ttctatatgc tgccactcct caattggatt agtctcatcc ttcaatgcta
     6781 tcatttcctt tgatattgga tcatctaaga aaccattatt atcatgacat taacctataa
     6841 aaataggcgt atcacgaggc ccttt
//
