LOCUS       sequence-380555-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy vector backbone with nourseothricin selectivity, encodes 
            for TDH3(pr) driven expression of mNeon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  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 7354)
  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-mNeon
FEATURES             Location/Qualifiers
     source          1..7354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      928..1516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1417..1436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1670..1687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1840..1845,1846..1863,1864..1870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1878..1894
                     /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     1883..1905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1902..1918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1937..1957
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1939..1957
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1982..2626
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2519..2542
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      3360..3607
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3376..3394)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3625..3644)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3626..3644)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3654..3671)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3654..3670)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3663..3685)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3811..4266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3898..3917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4108..4129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(4354..4551)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(5133..5476)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(5535..5554)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5714..5732)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5773..7119
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        7146..7250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7251..7319,7320..7354,1..757)
                     /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 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg cctttcgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctcagttt
     1981 atcattatca atactcgcca tttcaaagaa tacgtaaata attaatagta gtgattttcc
     2041 taactttatt tagtcaaaaa attagccttt taattctgct gtaacccgta catgcccaaa
     2101 atagggggcg ggttacacag aatatataac atcgtaggtg tctgggtgaa cagtttattc
     2161 ctggcatcca ctaaatataa tggagcccgc tttttaagct ggcatccaga aaaaaaaaga
     2221 atcccagcac caaaatattg ttttcttcac caaccatcag ttcataggtc cattctctta
     2281 gcgcaactac agagaacagg ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt
     2341 gatgcaacct gcctggagta aatgatgaca caaggcaatt gacccacgca tgtatctatc
     2401 tcattttctt acaccttcta ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa
     2461 aaggttgaaa ccagttccct gaaattattc ccctacttga ctaataagta tataaagacg
     2521 gtaggtattg attgtaattc tgtaaatcta tttcttaaac ttcttaaatt ctacttttat
     2581 agttagtctt ttttttagtt ttaaaacacc aagaacttag tttcgacgga tctagaacta
     2641 gtatggtctc aaaaggggag gaggacaaca tggcatcctt gccagcaact cacgaattac
     2701 atattttcgg aagtattaat ggtgttgact ttgatatggt tggacaaggc actggtaatc
     2761 caaacgatgg atacgaggaa cttaatctta aatcaacaaa aggcgatttg caattttccc
     2821 cctggattct tgtgccccat attgggtacg gctttcatca atatttacct taccctgatg
     2881 gaatgagtcc atttcaagcc gctatggtag acggttctgg ctaccaagtt catagaacca
     2941 tgcagtttga agacggcgct tccttaacag taaactacag gtacacgtac gagggttccc
     3001 atattaaggg agaagcccaa gttaagggta ctggttttcc ggctgatggt cccgttatga
     3061 ctaatagcct gacagctgct gactggtgta ggtctaaaaa gacatatcct aacgataaaa
     3121 caataatttc aacttttaag tggtcttata ccactggcaa tggtaaaaga tacagatcca
     3181 ctgcccgtac tacgtataca ttcgctaaac caatggccgc taattatctt aaaaatcaac
     3241 ccatgtacgt gtttaggaag actgaactaa aacatagtaa gaccgagttg aactttaagg
     3301 aatggcaaaa agccttcacc gatgtcatgg gaatggatga attatataag taactcgagt
     3361 catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga
     3421 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt
     3481 agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta
     3541 cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt
     3601 aatttgcggc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     3661 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3721 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3781 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     3841 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3901 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3961 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4021 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4081 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4141 accctatctc ggtctattct ttcgatttat aagggatttt gccgatttcg gcctattggt
     4201 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4261 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga
     4321 cactggatgg cggcgttagt atcgaatcga cagcagtata gcgaccagca ttcacatacg
     4381 attgacgcat gatattactt tctgcgcact taacttcgca tctgggcaga tgatgtcgag
     4441 gcgaaaaaaa atataaatca cgctaacatt tgattaaaat agaacaacta caatataaaa
     4501 aaactataca aatgacaagt tcttgaaaac aagaatcttt ttattgtcag tactgattat
     4561 caagggcaag gcattgacat atataatgcc tgttctccat ctgaagcagt tccgtcatac
     4621 aaggctgtat caaggccaca taaagtaaaa cccattcttc tgtatgcgtg gattgccggt
     4681 gcatttacgt tggtgacttc caaccacaaa tgacctgcac ctctttcacg tgcaaattct
     4741 gttgcaagtc ccatcagagc cctacccaca ccgtgacccc tatgttcagg agcaacttca
     4801 atgtcttcga ctgtaagtct tctgttccac cccgaatatg agactactac gaaaccagct
     4861 aaatctccgt cgtcgccgta ggccacaaat gttctactat ccgggtctcc gtcttcacca
     4921 tcgtcacttt catcatcgct ttcatcatca ggaaacactt tggttagagg tggatcgact
     4981 ggaacttctc ttaatgtaaa gccgtctcct gtagctgtaa cacggaaaac ggtatccgtc
     5041 gtgaaagaac catcaagagc ttcaatagct tctgcatcac ctgggacaga tgtacgatac
     5101 ctgtaggcgg tgtcgtctaa tgtagtaccc atggttgttt atgttcggat gtgatgtgag
     5161 aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat
     5221 cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac gcgtctgtga
     5281 ggggagcgtt tccctgctcg caggtctgca gcgaggagcc gtaatttttg cttcgcgccg
     5341 tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg ggctaaatgt
     5401 acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca aggagggtat
     5461 tctgggcctc catgtcgctg gccgggtgac ccggcgggga cgaggcaagc taaggtggta
     5521 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5581 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5641 gctgtgaccg tctccgggag ctgcatgcgt cagaggtttt caccgtcatc accgaaacgc
     5701 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5761 gtttcttagt atgatccaat atcaaaggaa atgatagcat tgaaggatga gactaatcca
     5821 attgaggagt ggcagcatat agaacagcta aagggtagtg ctgaaggaag catacgatac
     5881 cccgcatgga atgggataat atcacaggag gtactagact acctttcatc ctacataaat
     5941 agacgcatat aagtacgcat ttaagcataa acacgcacta tgccgttctt ctcatgtata
     6001 tatatataca ggcaacacgc agatataggt gcgacgtgaa cagtgagctg catgtgcgca
     6061 gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg ctttgaagtt cctattccga
     6121 agttcctatt ctctagctag aaagtatagg aacttcagag cgcttttgaa aaccaaaagc
     6181 gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa
     6241 tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta tatatctctg
     6301 tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg catctaaact
     6361 cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa aaattgtagt
     6421 aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt cctatacgta
     6481 gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga agaatcatca
     6541 acgctatcac cttctgttca caaagtatgc gcaatccaca tcggtataga atataatcgg
     6601 ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt aaacgcggga
     6661 agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc ttctcctaac
     6721 cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc gcacaaagga
     6781 gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat gcatttttgt
     6841 agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg ttgcatttct
     6901 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     6961 tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca
     7021 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
     7081 gcatttttgt tctacaaaat gaagcacaga tgcttcgttc aggtggcact tttcggggaa
     7141 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7201 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7261 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7321 acccagaaac gctggtgaaa gtaaaagatg ctga
//
