LOCUS       sequence-178967-        4787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4787)
  AUTHORS   Kistler AL, Guthrie C
  TITLE     Deletion of MUD2, the yeast homolog of U2AF65, can bypass the 
            requirement for sub2, an essential spliceosomal ATPase.
  JOURNAL   Genes Dev. 2001 Jan 1;15(1):42-9.
  PUBMED    11156604
REFERENCE   2  (bases 1 to 4787)
  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: "Genes 
            Dev."; date: "2001-01-1"; volume: "15(1)"; pages: "42-9"
COMMENT     Sequence Label: Sub2 in pRS314
FEATURES             Location/Qualifiers
     source          1..4787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     353..370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    487..508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(523..528,529..546,547..553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    561..577
                     /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)."
     primer_bind     566..588
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     585..601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     585..601
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        622..640
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    652..759
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     668..684
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     669..685
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(719..735)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(719..735)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(767..786)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(768..786)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(793..810)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(793..809)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..824)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      950..1405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1037..1056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1247..1268
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1508..2182)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2183..2463)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2484..2503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2603..2625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2663..2681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2722..3225
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3262..3366
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3367..3435,3436..4227)
                     /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(3585..3604)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4398..4787,1..199)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
       61 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      301 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      361 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      421 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      481 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      541 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      601 ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggtaccggg
      661 ccccccctcg aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga
      721 tccactagtt ctagagcggc cgccaccgcg gtggagctcc aattcgccct atagtgagtc
      781 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      841 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      901 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt
      961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1021 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1081 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1141 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1201 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1261 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1321 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1381 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     1441 gtatttcaca ccgcatagat cggcaagtgc acaaacaata cttaaataaa tactactcag
     1501 taataaccta tttcttagca tttttgacga aatttgctat tttgttagag tcttttacac
     1561 catttgtctc cacacctccg cttacatcaa caccaataac gccatttaat ctaagcgcat
     1621 caccaacatt ttctggcgtc agtccaccag ctaacataaa atgtaagctt tcggggctct
     1681 cttgccttcc aacccagtca gaaatcgagt tccaatccaa aagttcacct gtcccacctg
     1741 cttctgaatc aaacaaggga ataaacgaat gaggtttctg tgaagctgca ctgagtagta
     1801 tgttgcagtc ttttggaaat acgagtcttt taataactgg caaaccgagg aactcttggt
     1861 attcttgcca cgactcatct ccatgcagtt ggacgatatc aatgccgtaa tcattgacca
     1921 gagccaaaac atcctcctta ggttgattac gaaacacgcc aaccaagtat ttcggagtgc
     1981 ctgaactatt tttatatgct tttacaagac ttgaaatttt ccttgcaata accgggtcaa
     2041 ttgttctctt tctattgggc acacatataa tacccagcaa gtcagcatcg gaatctagag
     2101 cacattctgc ggcctctgtg ctctgcaagc cgcaaacttt caccaatgga ccagaactac
     2161 ctgtgaaatt aataacagac atactccaag ctgcctttgt gtgcttaatc acgtatactc
     2221 acgtgctcaa tagtcaccaa tgccctccct cttggccctc tccttttctt ttttcgaccg
     2281 aattaattct taatcggcaa aaaaagaaaa gctccggatc aagattgtac gtaaggtgac
     2341 aagctatttt tcaataaaga atatcttcca ctactgccat ctggcgtcat aactgcaaag
     2401 tacacatata ttacgatgct gttctattaa atgcttccta tattatatat atagtaatgt
     2461 cgtgatctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     2521 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     2581 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     2641 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     2701 ataataatgg tttcttagac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
     2761 aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg
     2821 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
     2881 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
     2941 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
     3001 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
     3061 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg
     3121 aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta atttatatat
     3181 ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt
     3241 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3301 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3361 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3421 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3481 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3541 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3601 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3661 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3721 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3781 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3841 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3901 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3961 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4021 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4081 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4141 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4201 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtga
//
