LOCUS       sequence-380565-        4980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy vector backbone with nourseothricin selectivity, contains 
            MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4980)
  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 4980)
  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
FEATURES             Location/Qualifiers
     source          1..4980
                     /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"
     CDS             1914..2294
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1937..1957
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1939..1957
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    1970..2077
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1994..2010
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1994..2010
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2044..2060)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2045..2061)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2085..2104)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2086..2104)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2114..2131)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2114..2130)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2123..2145)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2271..2726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2358..2377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2568..2589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(2813..3010)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(3592..3935)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(3994..4013)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4113..4135
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4173..4191)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4232..4735
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4772..4876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4877..4945,4946..4980,1..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 ccttttgctg
     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 agctccaccg
     1981 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg atatcaagct
     2041 tatcgatacc gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
     2101 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2161 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2221 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     2281 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2341 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2461 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2521 gtagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2581 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2641 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2701 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgg
     2761 gtatttcaca ccgcatagac actggatggc ggcgttagta tcgaatcgac agcagtatag
     2821 cgaccagcat tcacatacga ttgacgcatg atattacttt ctgcgcactt aacttcgcat
     2881 ctgggcagat gatgtcgagg cgaaaaaaaa tataaatcac gctaacattt gattaaaata
     2941 gaacaactac aatataaaaa aactatacaa atgacaagtt cttgaaaaca agaatctttt
     3001 tattgtcagt actgattatc aagggcaagg cattgacata tataatgcct gttctccatc
     3061 tgaagcagtt ccgtcataca aggctgtatc aaggccacat aaagtaaaac ccattcttct
     3121 gtatgcgtgg attgccggtg catttacgtt ggtgacttcc aaccacaagt gacctgcacc
     3181 tctttcacgt gcaaattctg ttgcaagtcc catcagagcc ctacccacac cgtgacccct
     3241 atgttcagga gcaacttcaa tgtcttcgac tgtaagtctt ctgttccacc ccgaatatga
     3301 gactactacg aaaccagcta aatctccgtc gtcgccgtag gccacaaatg ttctactatc
     3361 cgggtctccg tcttcaccat cgtcactttc atcatcgctt tcatcatcag gaaacacttt
     3421 ggttagaggt ggatcgactg gaacttctct taatgtaaag ccgtctcctg tagctgtaac
     3481 acggaaaacg gtatccgtcg tgaaagaacc atcaagagct tcaatagctt ctgcatcacc
     3541 tgggacagat gtacgatacc tgtaggcggt gtcgtctaat gtagtaccca tggttgttta
     3601 tgttcggatg tgatgtgaga actgtatcct agcaagattt taaaaggaag tatatgaaag
     3661 aagaacctca gtggcaaatc ctaacctttt atatttctct acaggggcgc ggcgtgggga
     3721 caattcaacg cgtctgtgag gggagcgttt ccctgctcgc aggtctgcag cgaggagccg
     3781 taatttttgc ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga ttatacatgg
     3841 ggatgtatgg gctaaatgta cgggcgacag tcacatcatg cccctgagct gcgcacgtca
     3901 agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc cggcggggac
     3961 gaggcaagct aaggtggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4021 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4081 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4141 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4201 taatgtcatg ataataatgg tttcttagac ggatcgcttg cctgtaactt acacgcgcct
     4261 cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta tttttatgtt
     4321 ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa
     4381 aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta
     4441 gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc
     4501 acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc
     4561 tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctttt
     4621 acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt tctattttta
     4681 atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac gtgatgaaaa
     4741 ggacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
//
