LOCUS       sequence-371398-        5988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  psac212-crispey-bar-ade2KO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5988)
  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 5988)
  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: pSAC212
FEATURES             Location/Qualifiers
     source          1..5988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(242..1045)
                     /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(1046..1261)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(1285..1304)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1404..1426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1464..1482)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1523..2026
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2063..2167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2168..2236,2237..3028)
                     /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(2386..2405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3199..3787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3688..3707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5458..5477)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5459..5477)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5487..5504)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5487..5503)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5496..5518)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(5644..5988,1..111)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5731..5750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5941..5962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
       61 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
      121 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat aactgatata
      181 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
      241 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
      301 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
      361 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
      421 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
      481 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
      541 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
      601 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
      661 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
      721 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
      781 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
      841 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
      901 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
      961 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     1021 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt tttgttctgt
     1081 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     1141 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     1201 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     1261 aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     1321 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1381 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1441 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1501 gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
     1561 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
     1621 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
     1681 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
     1741 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
     1801 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
     1861 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     1921 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     1981 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
     2041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2161 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2281 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2341 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2461 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2581 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2701 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2761 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3061 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3121 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3181 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3241 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3301 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3361 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3421 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3481 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3541 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3601 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3661 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3721 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3781 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3841 tggccttttg ctcaagtaga gggggtaatt tttccccttt attttgttca tacattctta
     3901 aattgctttg cctctccttt tggaaagcta tacttcggag cactgttgag cgaaggctca
     3961 ttagatatat tttctgtcat tttccttaac ccaaaaataa gggaaagggt ccaaaaagcg
     4021 ctcggacaac tgttgaccgt gatccgaagg actggctata cagtgttcac aaaatagcca
     4081 agctgaaaat aatgtgtagc tatgttcagt tagtttggct agcaaagata taaaagcagg
     4141 tcggaaatat ttatgggcat tattatgcag agcatcaaca tgataaaaaa aaacagttga
     4201 atattccctc aaaagaggtg cttgtagata acctccacga tggtgcacct tgggcaacac
     4261 aaaagtggca aatcatctac aatgcgcacc cttagcgaga ggtttatcat taaggtcaac
     4321 ctctggatgt tgtttcggca tcctgcattg aatctgagtt actgtctgtt tgaactgcag
     4381 ccaaagatgc gtgccgttgc agttagctaa caggccatgc tagcatcgat gcatgcacct
     4441 ggcgttcggc gatcgccata agagatctgc caattttaaa tttaaacccg tttcttctga
     4501 cgtaagggtg cgcagttgca gttagctaac aaccgtttca gagctatgct ggaaacagca
     4561 tagcaagttg aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     4621 tttttgatgg ccggcatggt cccagcctcc tcgctggcgc cggctgggca acaccttcgg
     4681 gtggcgaatg ggacttatgc gcacccttag cgagaggttt atcattaagg tcaacctctg
     4741 gatgttgttt cggcatcctg cattgaatct gagttactgt ctgttttcct ggcgcgccac
     4801 aggggaattg cttattaacg aaattgcctg aaggcctcac aactctggac attataccat
     4861 tgatgcttgc gtcacttcag gaaacccgtt tcttctgacg taagggtgcg caattaacga
     4921 aattgcccca gtttcagagc tatgctggaa acagcatagc aagttgaaat aaggctagtc
     4981 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tgacactgag tgagaaacgt
     5041 ccccgtcgta gtgtcggtaa tgcgttgttt caacgtagcc aattctcaca aagaaagtgg
     5101 aatattcatt catatcatat tttttctatt aactgcctgg tttcttttaa attttttatt
     5161 ggttgtcgac ttgaacggag tgacaatata tatatatata tttaataatg acatcattat
     5221 ctgtaaatct gattcttaat gctattctag ttatgtaaga gtggtccttt ccataaaaaa
     5281 aaaaaaaaga aaaaagaatt ttaggaatac aatgcagctt gtaagtaaaa tctggaatat
     5341 tcatatcgcc acaacttctt atgcttataa aagcactaat gcctgaattt atgttgaaaa
     5401 tatgtgtcac aaataaagaa actgtgacat ctgacacatt tccacgtacc caattcgccc
     5461 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     5521 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5581 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5641 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5701 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5821 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5941 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtct
//
