LOCUS       sequence-371542-        5484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  psac200-crispey-bar-vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5484)
  AUTHORS   Chen SA, Kern AF, Ang RML, Xie Y, Fraser HB
  TITLE     Gene-by-environment interactions are pervasive among natural genetic
            variants.
  JOURNAL   Cell Genom. 2023 Feb 28;3(4):100273. doi: 
            10.1016/j.xgen.2023.100273. eCollection 2023 Apr 12.
  PUBMED    37082145
REFERENCE   2  (bases 1 to 5484)
  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: "Cell 
            Genom."; date: "2023-02-28"; volume: "3(4):100273. doi"; pages: " 
            10.1016/j.xgen.2023.100273. eCollection 2023 Apr 12"
COMMENT     Sequence Label: pSAC200
FEATURES             Location/Qualifiers
     source          1..5484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    25..528
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        565..669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(670..738,739..1530)
                     /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     complement(888..907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1701..2289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2190..2209
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3456..3475)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3457..3475)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3485..3502)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3485..3501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3494..3516)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3642..4097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3729..3748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3939..3960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4228..5031)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5032..5247)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5271..5290)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5390..5412
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5450..5468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct
       61 tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt
      121 tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa
      181 acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga
      241 aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat
      301 tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc
      361 aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt
      421 cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata
      481 tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccca
      541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1381 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1681 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1741 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1801 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1861 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1921 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1981 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2041 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2101 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2161 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2221 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2281 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2341 gctggccttt tgctcaagta gagggggtaa tttttcccct ttattttgtt catacattct
     2401 taaattgctt tgcctctcct tttggaaagc tatacttcgg agcactgttg agcgaaggct
     2461 cattagatat attttctgtc attttcctta acccaaaaat aagggaaagg gtccaaaaag
     2521 cgctcggaca actgttgacc gtgatccgaa ggactggcta tacagtgttc acaaaatagc
     2581 caagctgaaa ataatgtgta gctatgttca gttagtttgg ctagcaaaga tataaaagca
     2641 ggtcggaaat atttatgggc attattatgc agagcatcaa catgataaaa aaaaacagtt
     2701 gaatattccc tcaaaagagg tgcttgtaga taacctccac gatggtgcac cttgggcaac
     2761 acaaaagtgg caaatcatct acaatgcgca cccttagcga gaggtttatc attaaggtca
     2821 acctctggat gttgtttcgg catcctgcat tgaatctgag ttactgtctg tttgaactgc
     2881 agccaaagat gcgtgccgtt gcagttagct aacaggccat gcggccgcgt ttcagagcta
     2941 tgctggaaac agcatagcaa gttgaaataa ggctagtccg ttatcaactt gaaaaagtgg
     3001 caccgagtcg gtgctttttg acactgagtg agaaacgtcc ccgtcgtagt gtcggtaatg
     3061 cgttgtttca acgtagccaa ttctcacaaa gaaagtggaa tattcattca tatcatattt
     3121 tttctattaa ctgcctggtt tcttttaaat tttttattgg ttgtcgactt gaacggagtg
     3181 acaatatata tatatatatt taataatgac atcattatct gtaaatctga ttcttaatgc
     3241 tattctagtt atgtaagagt ggtcctttcc ataaaaaaaa aaaaaagaaa aaagaatttt
     3301 aggaatacaa tgcagcttgt aagtaaaatc tggaatattc atatcgccac aacttcttat
     3361 gcttataaaa gcactaatgc ctgaatttat gttgaaaata tgtgtcacaa ataaagaaac
     3421 tgtgacatct gacacatttc cacgtaccca attcgcccta tagtgagtcg tattacgcgc
     3481 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     3541 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     3601 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     3661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3841 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3901 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3961 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4021 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4081 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4141 caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     4201 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     4261 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     4321 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     4381 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     4441 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     4501 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     4561 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     4621 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     4681 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     4741 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     4801 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     4861 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     4921 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     4981 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     5041 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     5101 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     5161 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     5221 agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc
     5281 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5341 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5401 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     5461 cgcctatttt tataggttaa tgtc
//
