LOCUS       sequence-380557-        5860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low 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 5860)
  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 5860)
  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: pRSII41N-TDH3pr
FEATURES             Location/Qualifiers
     source          1..5860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        72..415
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      997..1194
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(1281..1736)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1418..1439)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1630..1649
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1862..1884
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1876..1893
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1877..1893
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1903..1922
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1903..1921
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1940..2187)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2153..2171
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     2189..2205
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2190..2206
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2240..2256)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2240..2256)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2261..2905)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(2345..2368)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(2930..2950)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2930..2948)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2969..2985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2969..2985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2982..3004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2993..3009
                     /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(3017..3023,3024..3041,3042..3047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3062..3083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3200..3217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3371..3959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3451..3470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4130..4921,4922..4990))
                     /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     4753..4772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4991..5095)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5132..5635
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5676..5694
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5732..5754)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(5854..5860,1..13)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agagcagatt gtactgagag tgcaccatac caccttagct tgcctcgtcc ccgccgggtc
       61 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
      121 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
      181 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
      241 agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc
      301 ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc
      361 cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg
      421 tactacatta gacgacaccg cctacaggta tcgtacatct gtcccaggtg atgcagaagc
      481 tattgaagct cttgatggtt ctttcacgac ggataccgtt ttccgtgtta cagctacagg
      541 agacggcttt acattaagag aagttccagt cgatccacct ctaaccaaag tgtttcctga
      601 tgatgaaagc gatgatgaaa gtgacgatgg tgaagacgga gacccggata gtagaacatt
      661 tgtggcctac ggcgacgacg gagatttagc tggtttcgta gtagtctcat attcggggtg
      721 gaacagaaga cttacagtcg aagacattga agttgctcct gaacataggg gtcacggtgt
      781 gggtagggct ctgatgggac ttgcaacaga atttgcacgt gaaagaggtg caggtcactt
      841 gtggttggaa gtcaccaacg taaatgcacc ggcaatccac gcatacagaa gaatgggttt
      901 tactttatgt ggccttgata cagccttgta tgacggaact gcttcagatg gagaacaggc
      961 attatatatg tcaatgcctt gcccttgata atcagtactg acaataaaaa gattcttgtt
     1021 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     1081 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     1141 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     1201 attcgatact aacgccgcca tccagtgtct atgcggtgtg aaatacccac agatgcgtaa
     1261 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1321 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     1381 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     1441 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctaccagg gcgatggccc
     1501 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     1561 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     1621 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     1681 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     1741 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     1801 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1861 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     1921 ggcgaattgg gtaccggccg caaattaaag ccttcgagcg tcccaaaacc ttctcaagca
     1981 aggttttcag tataatgtta catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt
     2041 gaaatataaa taacgttctt aatactaaca taactataaa aaaataaata gggacctaga
     2101 cttcaggttg tctaactcct tccttttcgg ttagagcgga tgtgggggga gggcgtgaat
     2161 gtaagcgtga cataactaat tacatgactc gaggtcgacg gtatcgataa gcttgatatc
     2221 gaattcctgc agcccggggg atccactagt tctagatccg tcgaaactaa gttcttggtg
     2281 ttttaaaact aaaaaaaaga ctaactataa aagtagaatt taagaagttt aagaaataga
     2341 tttacagaat tacaatcaat acctaccgtc tttatatact tattagtcaa gtaggggaat
     2401 aatttcaggg aactggtttc aacctttttt ttcagctttt tccaaatcag agagagcaga
     2461 aggtaataga aggtgtaaga aaatgagata gatacatgcg tgggtcaatt gccttgtgtc
     2521 atcatttact ccaggcaggt tgcatcactc cattgaggtt gtgcccgttt tttgcctgtt
     2581 tgtgcccctg ttctctgtag ttgcgctaag agaatggacc tatgaactga tggttggtga
     2641 agaaaacaat attttggtgc tgggattctt tttttttctg gatgccagct taaaaagcgg
     2701 gctccattat atttagtgga tgccaggaat aaactgttca cccagacacc tacgatgtta
     2761 tatattctgt gtaacccgcc ccctattttg ggcatgtacg ggttacagca gaattaaaag
     2821 gctaattttt tgactaaata aagttaggaa aatcactact attaattatt tacgtattct
     2881 ttgaaatggc gagtattgat aatgataaac tgagctccag cttttgttcc ctttagtgag
     2941 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3001 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3061 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3121 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3181 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3241 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3301 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3361 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3421 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3481 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3541 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3601 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3661 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3721 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3781 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3841 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3901 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3961 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4021 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4081 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5101 ttccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt
     5161 aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa
     5221 aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac
     5281 cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg
     5341 tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta
     5401 tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg
     5461 aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt
     5521 atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc
     5581 aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtct
     5641 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5701 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5761 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5821 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
//
