LOCUS       sequence-380544-        6695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy vector backbone with nourseothricin selectivity, contains 
            TDH3(pr)-MCS-CYC1(3'UTR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  AUTHORS   Smith JR, Sislak CD, Fernandez Mendoza P, Carmichael L, Lewis AG, 
            Chen A, Jiang GZ, Gibney PA
  TITLE     Auxotrophy-Independent Plasmid Shuttle Vectors for Applications in 
            Diverse Yeasts
  JOURNAL   Appl Microbiol. 2024; 4(1):453-469. doi: 
            10.3390/applmicrobiol4010031
REFERENCE   2  (bases 1 to 6695)
  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: "Appl 
            Microbiol."; date: "2024"; volume: " 4(1):453-469. doi"; pages: " 
            10.3390/applmicrobiol401003"
COMMENT     Sequence Label: pRSII42N-TDH3pr
FEATURES             Location/Qualifiers
     source          1..6695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     123..142
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        201..544
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1126..1323
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(1411..1866)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1548..1569)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1760..1779
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1992..2014
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2006..2023
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2007..2023
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2033..2052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2033..2051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2070..2318)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2284..2302
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     2320..2336
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2321..2337
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2371..2387)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2371..2387)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2392..3036)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(2476..2499)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(3061..3081)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3061..3079)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3100..3116)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3100..3116)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3113..3135)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3124..3140
                     /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(3148..3154,3155..3172,3173..3178))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3193..3214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3331..3348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3502..4090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3582..3601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4261..5052,5053..5121))
                     /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     4884..4903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5122..5226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5253..6599
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6640..6658
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctctgacgc atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
       61 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
      121 tgcggcatca gagcagattg tactgagagt gcaccatacc accttagctt gcctcgtccc
      181 cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg
      241 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
      301 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
      361 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
      421 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
      481 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
      541 aaccatgggt actacattag acgacaccgc ctacaggtat cgtacatctg tcccaggtga
      601 tgcagaagct attgaagctc ttgatggttc tttcacgacg gataccgttt tccgtgttac
      661 agctacagga gacggcttta cattaagaga agttccagtc gatccacctc taaccaaagt
      721 gtttcctgat gatgaaagcg atgatgaaag tgacgatggt gaagacggag acccggatag
      781 tagaacattt gtggcctacg gcgacgacgg agatttagct ggtttcgtag tagtctcata
      841 ttcggggtgg aacagaagac ttacagtcga agacattgaa gttgctcctg aacatagggg
      901 tcacggtgtg ggtagggctc tgatgggact tgcaacagaa tttgcacgtg aaagaggtgc
      961 aggtcatttg tggttggaag tcaccaacgt aaatgcaccg gcaatccacg catacagaag
     1021 aatgggtttt actttatgtg gccttgatac agccttgtat gacggaactg cttcagatgg
     1081 agaacaggca ttatatatgt caatgccttg cccttgataa tcagtactga caataaaaag
     1141 attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt
     1201 taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg
     1261 aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata
     1321 ctgctgtcga ttcgatacta acgccgccat ccagtgtcta tgcggtgtga aataccgcac
     1381 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat
     1441 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1501 tcccttataa atcgaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1561 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1621 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1681 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1741 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1801 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1861 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1921 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1981 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     2041 ctcactatag ggcgaattgg gtaccggccg caaattaaag ccttcgagcg tcccaaaacc
     2101 ttctcaagca aggttttcag tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa
     2161 agaaaaattt gaaatataaa taacgttctt aatactaaca taactataaa aaaataaata
     2221 gggacctaga cttcaggttg tctaactcct tccttttcgg ttagagcgga tgtggggggg
     2281 agggcgtgaa tgtaagcgtg acataactaa ttacatgact cgaggtcgac ggtatcgata
     2341 agcttgatat cgaattcctg cagcccgggg gatccactag ttctagatcc gtcgaaacta
     2401 agttcttggt gttttaaaac taaaaaaaag actaactata aaagtagaat ttaagaagtt
     2461 taagaaatag atttacagaa ttacaatcaa tacctaccgt ctttatatac ttattagtca
     2521 agtaggggaa taatttcagg gaactggttt caaccttttt tttcagcttt ttccaaatca
     2581 gagagagcag aaggtaatag aaggtgtaag aaaatgagat agatacatgc gtgggtcaat
     2641 tgccttgtgt catcatttac tccaggcagg ttgcatcact ccattgaggt tgtgcccgtt
     2701 ttttgcctgt ttgtgcccct gttctctgta gttgcgctaa gagaatggac ctatgaactg
     2761 atggttggtg aagaaaacaa tattttggtg ctgggattct ttttttttct ggatgccagc
     2821 ttaaaaagcg ggctccatta tatttagtgg atgccaggaa taaactgttc acccagacac
     2881 ctacgatgtt atatattctg tgtaacccgc cccctatttt gggcatgtac gggttacagc
     2941 agaattaaaa ggctaatttt ttgactaaat aaagttagga aaatcactac tattaattat
     3001 ttacgtattc tttgaaatgg cgagtattga taatgataaa ctgagctcca gcttttgttc
     3061 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3121 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3181 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3241 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3301 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3361 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3421 aggggataac gcaggaaaga acatgtgagc aaaaggccag cgaaaggcca ggaaccgtaa
     3481 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3541 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3601 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3661 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3721 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3781 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3841 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3901 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3961 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4021 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4081 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4141 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4201 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4261 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4321 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4381 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4441 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4501 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4561 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4621 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4681 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4741 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4801 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4861 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4921 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4981 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5041 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5101 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5161 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5221 tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag
     5281 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5341 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     5401 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     5461 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     5521 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     5581 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     5641 ggtccgttaa ggttaggaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     5701 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     5761 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     5821 ggtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5881 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5941 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     6001 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     6061 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     6121 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     6181 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga
     6241 gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat
     6301 gcaacgcgag ctgcgcacat gcagctcact gttcacgtcg cacctatatc tgcgtgttgc
     6361 ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact
     6421 tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca
     6481 ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc
     6541 cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca
     6601 tactaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6661 tttcgtctcg cgcgtttcgg tgatgacggt gaaaa
//
