LOCUS       sequence-380549-        6521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low 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 6521)
  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 6521)
  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-mNeon
FEATURES             Location/Qualifiers
     source          1..6521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(103..122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      916..1504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1405..1424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1658..1675
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1792..1813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1828..1833,1834..1851,1852..1858)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1866..1882
                     /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     1871..1893
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1890..1906
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1890..1906
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1925..1945
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1927..1945
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1970..2614
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2507..2530
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      3349..3596
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3365..3383)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3614..3633)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3615..3633)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3643..3660)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3643..3659)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3652..3674)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3800..4255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3887..3906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4097..4118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(4342..4539)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(5121..5464)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(5523..5542)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5642..5664
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5702..5720)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5761..6264
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6301..6405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6406..6474,6475..6521,1..745)
                     /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 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
       61 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      121 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      181 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      241 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      301 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      361 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      421 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      481 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      541 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      601 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      661 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      721 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
      781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1681 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1741 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     1801 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1861 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1921 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctcagtttat cattatcaat
     1981 actcgccatt tcaaagaata cgtaaataat taatagtagt gattttccta actttattta
     2041 gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg
     2101 ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact
     2161 aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca
     2221 aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag
     2281 agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc
     2341 ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac
     2401 accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc
     2461 agttccctga aattattccc ctacttgact aataagtata taaagacggt aggtattgat
     2521 tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt
     2581 ttttagtttt aaaacaccaa gaacttagtt tcgacggatt ctagaactag tatggtctca
     2641 aaaggggagg aggacaacat ggcatccttg ccagcaactc acgaattaca tattttcgga
     2701 agtattaatg gtgttgactt tgatatggtt ggacaaggca ctggtaatcc aaacgatgga
     2761 tacgaggaac ttaatcttaa atcaacaaaa ggcgatttgc aattttcccc ctggattctt
     2821 gtgccccata ttgggtacgg ctttcatcaa tatttacctt accctgatgg aatgagtcca
     2881 tttcaagccg ctatggtaga cggttctggc taccaagttc atagaaccat gcagtttgaa
     2941 gacggcgctt ccttaacagt aaactacagg tacacgtacg agggttccca tattaaggga
     3001 gaagcccaag ttaagggtac tggttttccg gctgatggtc ccgttatgac taatagcctg
     3061 acagctgctg actggtgtag gtctaaaaag acatatccta acgataaaac aataatttca
     3121 acttttaagt ggtcttatac cactggcaat ggtaaaagat acagatccac tgcccgtact
     3181 acgtatacat tcgctaaacc aatggctgcc aattatctta aaaatcaacc catgtacgtg
     3241 tttaggaaga ctgaactaaa acatagtaag accgagttga actttaagga atggcaaaaa
     3301 gccttcaccg atgtcatggg aatggatgaa ttatataagt aactcgagtc atgtaattag
     3361 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     3421 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     3481 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     3541 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc
     3601 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
     3661 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     3721 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     3781 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3841 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3901 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3961 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4021 gatggttcac gtagtgggcc atcgccctgg tagacggttt ttcgcccttt gacgttggag
     4081 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4141 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4201 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     4261 tgcggtattt tctccttacg catctgtggg tatttcacac cgcatagaca ctggatggcg
     4321 gcgttagtat cgaatcgaca gcagtatagc gaccagcatt cacatacgat tgacgcatga
     4381 tattactttc tgcgcactta acttcgcatc tgggcagatg atgtcgaggc gaaaaaaaat
     4441 ataaatcacg ctaacatttg attaaaatag aacaactaca atataaaaaa actatacaaa
     4501 tgacaagttc ttgaaaacaa gaatcttttt attgtcagta ctgattatca agggcaaggc
     4561 attgacatat ataatgcctg ttctccatct gaagcagttc cgtcatacaa ggctgtatca
     4621 aggccacata aagtaaaacc cattcttctg tatgcgtgga ttgccggtgc atttacgttg
     4681 gtgacttcca accacaagtg acctgcacct ctttcacgtg caaattctgt tgcaagtccc
     4741 atcagagccc tacccacacc gtgaccccta tgttcaggag caacttcaat gtcttcgact
     4801 gtaagtcttc tgttccaccc cgaatatgag actactacga aaccagctaa atctccgtcg
     4861 tcgccgtagg ccacaaatgt tctactatcc gggtctccgt cttcaccatc gtcactttca
     4921 tcatcgcttt catcatcagg aaacactttg gttagaggtg gatcgactgg aacttctctt
     4981 aatgtaaagc cgtctcctgt agctgtaaca cggaaaacgg tatccgtcgt gaaagaacca
     5041 tcaagagctt caatagcttc tgcatcacct gggacagatg tacgatacct gtaggcggtg
     5101 tcgtctaatg tagtacccat ggttgtttat gttcggatgt gatgtgagaa ctgtatccta
     5161 gcaagatttt aaaaggaagt atatgaaaga agaacctcag tggcaaatcc taacctttta
     5221 tatttctcta caggggcgcg gcgtggggac aattcaacgc gtctgtgagg ggagcgtttc
     5281 cctgctcgca ggtctgcagc gaggagccgt aatttttgct tcgcgccgtg cggccatcaa
     5341 aatgtatgga tgcaaatgat tatacatggg gatgtatggg ctaaatgtac gggcgacagt
     5401 cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag gagggtattc tgggcctcca
     5461 tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta aggtggtatg gtgcactctc
     5521 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5581 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5641 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5701 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5761 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
     5821 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
     5881 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
     5941 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
     6001 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
     6061 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
     6121 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
     6181 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
     6241 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
     6301 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6361 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6421 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6481 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg g
//
