LOCUS       sequence-176075-        6035 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6035)
  AUTHORS   Fang F, Salmon K, Shen MW, Aeling KA, Ito E, Irwin B, Tran UP, 
            Hatfield GW, Da Silva NA, Sandmeyer S
  TITLE     A vector set for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2011 Feb;28(2):123-36. doi: 10.1002/yea.1824. Epub 2010 Oct 
            8.
  PUBMED    20936606
REFERENCE   2  (bases 1 to 6035)
  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"; doi: "10.1002/yea.1824"; 
            journalName: "Yeast"; date: "2011-02"; volume: "28"; issue: "2"; 
            pages: "123-36"
COMMENT     Sequence Label: pXP420
FEATURES             Location/Qualifiers
     source          1..6035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      451..698
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(467..485)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     861..879
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(917..939)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1039..1058
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1252..1274
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1266..1283
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1267..1283
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1328..1361)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1486..1673
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1674..2336
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     protein_bind    complement(2538..2571)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2594..3936
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3557..3604)
                     /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(3955..3971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3955..3971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3968..3990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3979..3995
                     /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(4003..4009,4010..4027,4028..4033))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4048..4069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4186..4203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4357..4945)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4437..4456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5116..5907,5908..5976))
                     /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     5739..5758
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        join(6035,1..413)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc
       61 tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc
      121 cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa
      181 gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc
      241 tttttcttga aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt
      301 taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac
      361 aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct aagttttaat
      421 tacaaaacta gtgatatctg cgcactcgag tcatgtaatt agttatgtca cgcttacatt
      481 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
      541 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
      601 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
      661 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccaatattat tgaagcattt
      721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      781 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
      841 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
      901 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
      961 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1021 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     1081 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     1141 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     1201 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1261 gacgttgtaa aacgacggcc agtgccaagc ttgcatgcct gcaggtcgac tctagaggat
     1321 ccccgggata acttcgtata gcatacatta tacgaagtta tcgttttaag agcttggtga
     1381 gcgctaggag tcactgccag gtatcgtttg aacacggcat tagtcaggga agtcataaca
     1441 cagtcctttc ccgcaatttt ctttttctat tactcttggc ctcctctagt acactctata
     1501 tttttttatg cctcggtaat gattttcatt tttttttttc cacctagcgg atgactcttt
     1561 ttttttctta gcgattggca ttatcacata atgaattata cattatataa agtaatgtga
     1621 tttcttcgaa gaatatacta aaaaatgagc aggcaagata aacgaaggca aagatgacag
     1681 agcagaaagc cctagtaaag cgtattacaa atgaaaccaa gattcagatt gcgatctctt
     1741 taaagggtgg tcccctagcg atagagcact cgatcttccc agaaaaagag gcagaagcag
     1801 tagcagaaca ggccacacaa tcgcaagtga ttaacgtcca cacaggtata gggtttctgg
     1861 accatatgat acatgctctg gccaagcatt ccggctggtc gctaatcgtt gagtgcattg
     1921 gtgacttaca catagacgac catcacacca ctgaagactg cgggattgct ctcggtcaag
     1981 cttttaaaga ggccctaggg gccgtgcgtg gagtaaaaag gtttggatca ggatttgcgc
     2041 ctttggatga ggcactttcc agagcggtgg tagatctttc gaacaggccg tacgcagttg
     2101 tcgaacttgg tttgcaaagg gagaaagtag gagatctctc ttgcgagatg atcccgcatt
     2161 ttcttgaaag ctttgcagag gctagcagaa ttaccctcca cgttgattgt ctgcgaggca
     2221 agaatgatca tcaccgtagt gagagtgcgt tcaaggctct tgcggttgcc ataagagaag
     2281 ccacctcgcc caatggtacc aacgatgttc cctccaccaa aggtgttctt atgtagtgac
     2341 accgattatt taaagctgca gcatacgata tatatacatg tgtatatatg tatacctatg
     2401 aatgtcagta agtatgtata cgaacagtat gatactgaag atgacaaggt aatgcatcat
     2461 tctatacgtg tcattctgaa cgaggcgcgc tttccttttt tctttttgct ttttcttttt
     2521 ttttctcttg aactcgaata acttcgtata gcatacatta tacgaagtta tcccgggtac
     2581 cgagctcgaa ttcaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     2641 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     2701 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     2761 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2821 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     2881 gcatcccgag agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     2941 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     3001 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     3061 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     3121 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     3181 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     3241 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     3301 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     3361 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     3421 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     3481 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg
     3541 ttttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
     3601 cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca
     3661 gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa
     3721 gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga
     3781 tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct
     3841 tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc
     3901 atccttcaat gctatcattt cctttgatat tggatcatac gaattcgtaa tcatgtcata
     3961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4861 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4921 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4981 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5041 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5101 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5161 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5221 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5281 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5341 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5401 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5461 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5521 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5581 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5641 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5701 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5761 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5821 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5881 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5941 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattaccg
     6001 cgaatcctta catcacaccc aatcccccac aagtg
//
