LOCUS       sequence-380551-        6095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy vector backbone with G418 selectivity, contains 
            TDH3(pr)-MCS-CYC1(3'UTR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6095)
  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 6095)
  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: pRSII41K-TDH3pr
FEATURES             Location/Qualifiers
     source          1..6095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      919..1507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1408..1427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1661..1678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1795..1816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1831..1836,1837..1854,1855..1861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1869..1885
                     /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     1874..1896
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1893..1909
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1893..1909
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1928..1948
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1930..1948
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1973..2617
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2510..2533
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     2622..2638
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2622..2638
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2672..2688)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2673..2689)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      2691..2938
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2707..2725)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2956..2975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2957..2975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2985..3002)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2985..3001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2994..3016)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3142..3597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3229..3248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3439..3460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            complement(3685..5041)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(3685..3882)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(3888..4697)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4611..4630
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4698..5041)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(5100..5119)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5219..5241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5279..5297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5338..5841
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5878..5982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5983..6051,6052..6095,1..748)
                     /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"
ORIGIN
        1 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
       61 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccaggt tggactcaag
     1201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1681 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1741 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1801 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1861 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1921 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcagtt tatcattatc
     1981 aatactcgcc atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat
     2041 ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc
     2101 gggttacaca gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc
     2161 actaaatata atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca
     2221 ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta
     2281 cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc
     2341 tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct
     2401 tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa
     2461 accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt
     2521 gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct
     2581 tttttttagt tttaaaacac caagaactta gtttcgacgg atctagaact agtggatccc
     2641 ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctcgag tcatgtaatt
     2701 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     2761 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     2821 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     2881 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
     2941 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     3001 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3061 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3121 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3181 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3241 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3301 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3361 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3421 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3481 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3541 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     3601 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag acactggatg
     3661 gcggcgttag tatcgaatcg acagcagtat agcgaccagc attcacatac gattgacgca
     3721 tgatattact ttctgcgcac ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa
     3781 aatataaatc acgctaacat ttgattaaaa tagaacaact acaatataaa aaaactatac
     3841 aaatgacaag ttcttgaaaa caagaatctt tttattgtca gtactgatta gaaaaactca
     3901 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3961 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     4021 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     4081 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     4141 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     4201 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     4261 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     4321 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     4381 tggaatgctg ttttgccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     4441 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     4501 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     4561 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     4621 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgaaacgtga
     4681 gtcttttcct tacccatggt tgtttatgtt cggatgtgat gtgagaactg tatcctagca
     4741 agattttaaa aggaagtata tgaaagaaga acctcagtgg caaatcctaa ccttttatat
     4801 ttctctacag gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga gcgtttccct
     4861 gctcgcaggt ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat
     4921 gtatggatgc aaatgattat acatggggat gtatgggcta aatgtacggg cgacagtcac
     4981 atcatgcccc tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg gcctccatgt
     5041 cgctggccgg gtgacccggc ggggacgagg caagctaagg tggtatggtg cactctcagt
     5101 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5161 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5221 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5281 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacggat
     5341 cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat ttgggaattt
     5401 actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa ataaagaagg
     5461 tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa
     5521 agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga tatacattcg
     5581 attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc tacacagaca
     5641 agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa aggtagtatt
     5701 tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat tttttcttta
     5761 atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat ttaaattata
     5821 attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg aaatgtgcgc
     5881 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6001 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6061 acgctggtga aagtaaaaga tgctgaagat cagtt
//
