LOCUS       sequence-172992-        6028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector to facilitate gene expression for metabolic 
            engineering in S. cerevisiae. Contains TEF1 promoter and CYC1 
            terminator for insertion of gene of interest. Contains HIS3 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6028)
  TITLE     pXP420 repaired Amp promoter
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6028)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pXP420 v2 (repaired Amp promoter)
FEATURES             Location/Qualifiers
     source          1..6028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1181..1594
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      1632..1879
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1648..1666)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     1944..1962
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2000..2022)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2122..2141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2335..2357
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2349..2366
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2350..2366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2411..2444)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2569..2756
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2757..3419
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAAVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     protein_bind    complement(3621..3654)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      3677..5019
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4640..4687)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(5038..5054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5038..5054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5051..5073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5062..5078
                     /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(5086..5092,5093..5110,5111..5116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5131..5152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5269..5286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5440..6028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5520..5539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgaata
     1141 ttaccgcgaa tccttacatc acacccaatc ccccacaagt gatcccccac acaccatagc
     1201 ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc gcgcatcgcc
     1261 gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct tcctctaggg
     1321 tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt
     1381 ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg aaaatttttt
     1441 tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat aaacggtctt
     1501 caatttctca agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgc
     1561 tcattagaaa gaaagcatag caatctaatc taagttttaa ttacaaaact agtgatatct
     1621 gcgcactcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     1681 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1741 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     1801 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     1861 ctcgaaggct ttaatttgcg gccccccgaa aagtgccacc tgacgtctaa gaaaccatta
     1921 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     1981 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     2041 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     2101 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     2161 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     2221 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     2281 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     2341 cacgacgttg taaaacgacg gccagtgcca agcttgcatg cctgcaggtc gactctagag
     2401 gatccccggg ataacttcgt atagcataca ttatacgaag ttatcgtttt aagagcttgg
     2461 tgagcgctag gagtcactgc caggtatcgt ttgaacacgg cattagtcag ggaagtcata
     2521 acacagtcct ttcccgcaat tttctttttc tattactctt ggcctcctct agtacactct
     2581 atattttttt atgcctcggt aatgattttc attttttttt ttccacctag cggatgactc
     2641 tttttttttc ttagcgattg gcattatcac ataatgaatt atacattata taaagtaatg
     2701 tgatttcttc gaagaatata ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga
     2761 cagagcagaa agccctagta aagcgtatta caaatgaaac caagattcag attgcgatct
     2821 ctttaaaggg tggtccccta gcgatagagc actcgatctt cccagaaaaa gaggcagaag
     2881 cagtagcaga acaggccaca caatcgcaag tgattaacgt ccacacaggt atagggtttc
     2941 tggaccatat gatacatgct ctggccaagc attccggctg gtcgctaatc gttgagtgca
     3001 ttggtgactt acacatagac gaccatcaca ccactgaaga ctgcgggatt gctctcggtc
     3061 aagcttttaa agaggcccta ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg
     3121 cgcctttgga tgaggcactt tccagagcgg cggtagatct ttcgaacagg ccgtacgcag
     3181 ttgtcgaact tggtttgcaa agggagaaag taggagatct ctcttgcgag atgatcccgc
     3241 attttcttga aagctttgca gaggctagca gaattaccct ccacgttgat tgtctgcgag
     3301 gcaagaatga tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt gccataagag
     3361 aagccacctc gcccaatggt accaacgatg ttccctccac caaaggtgtt cttatgtagt
     3421 gacaccgatt atttaaagct gcagcatacg atatatatac atgtgtatat atgtatacct
     3481 atgaatgtca gtaagtatgt atacgaacag tatgatactg aagatgacaa ggtaatgcat
     3541 cattctatac gtgtcattct gaacgaggcg cgctttcctt ttttcttttt gctttttctt
     3601 tttttttctc ttgaactcga ataacttcgt atagcataca ttatacgaag ttatcccggg
     3661 taccgagctc gaattcaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     3721 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     3781 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     3841 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     3901 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     3961 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     4021 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     4081 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     4141 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     4201 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     4261 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     4321 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     4381 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     4441 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     4501 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     4561 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
     4621 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
     4681 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
     4741 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
     4801 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
     4861 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
     4921 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
     4981 ctcatccttc aatgctatca tttcctttga tattggatca tacgaattcg taatcatgtc
     5041 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5101 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5161 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5221 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5281 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5341 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5401 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5461 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5521 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5581 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5641 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5701 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5761 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5821 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5881 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5941 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6001 gattacgcgc agaaaaaaag gatctcaa
//
