LOCUS       sequence-12717-e        6005 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 6005)
  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 6005)
  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: pXP622
FEATURES             Location/Qualifiers
     source          1..6005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      598..845
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(614..632)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(891..924)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2366..2399)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2426..3768
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    2758..2805
                     /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(3784..3800)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3784..3800)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3797..3819)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3808..3824
                     /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(3832..3838,3839..3856,3857..3862))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3877..3898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4015..4032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4186..4774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4266..4285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4945..5736,5737..5805))
                     /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     5568..5587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5806..5910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5978..5996
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgct cgtaggaaca atttcgggcc cctgcgtgtt cttctgaggt tcatctttta
      241 catttgcttc tgctggataa ttttcagagg caacaaggaa aaattagatg gcaaaaagtc
      301 gtctttcaag gaaaaatccc caccatcttt cgagatcccc tgtaacttat tggcaactga
      361 aagaatgaaa aggaggaaaa tacaaaatat actagaactg aaaaaaaaaa agtataaata
      421 gagacgatat atgccaatac ttcacaatgt tcgaatctat tcttcatttg cagctattgt
      481 aaaataataa aacatcaaga acaaacaagc tcaacttgtc ttttctaaga acaaagaata
      541 aacacaaaaa caaaaagttt ttttaatttt aatcaaaaaa ctagtgatat cctcgagtca
      601 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
      661 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
      721 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
      781 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
      841 tttgcggcca agcttgcatg cctgcaggtc gactctagag gatccccggg ataacttcgt
      901 atagcataca ttatacgaag ttattaacca ttattttttt cctcaacata acgagaacac
      961 acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga
     1021 ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga
     1081 ggccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac
     1141 aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc
     1201 aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat atatatattt
     1261 caaggatata ccattctaat gtctgcccct aagaagatcg tcgttttgcc aggtgaccac
     1321 gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc tgatgttcgt
     1381 tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat cgaagctaca
     1441 ggtgtcccac ttccagatga ggcgctggaa gcctccaaga aggctgatgc cgttttgtta
     1501 ggtgctgtgg gtggtcctaa atggggtaca ggtagtgtta gacctgaaca aggtttacta
     1561 aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt tgcatccgac
     1621 tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga cttcgttgtt
     1681 gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagatga tggtgatggt
     1741 gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac aagaatggcc
     1801 gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga taaagctaat
     1861 gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa gaacgaattt
     1921 cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct agttaagaac
     1981 ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat catctccgat
     2041 gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg
     2101 ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc tccagatttg
     2161 ccaaagaata aggtcaaccc tatcgccact atcttgtctg ctgcaatgat gttgaaattg
     2221 tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa ggttttggat
     2281 gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt cggtgatgct
     2341 gtcgccgaag aagttaagaa aatccataac ttcgtatagc atacattata cgaagttatc
     2401 ccgggtaccg agctcgaatt cgtatgatcc aatatcaaag gaaatgatag cattgaagga
     2461 tgagactaat ccaattgagg agtggcagca tatagaacag ctaaagggta gtgctgaagg
     2521 aagcatacga taccccgcat ggaatgggat aatatcacag gaggtactag actacctttc
     2581 atcctacata aatagacgca tataagtacg catttaagca taaacacgca ctatgccgtt
     2641 cttctcatgt atatatatat acaggcaaca cgcagatata ggtgcgacgt gaacagtgag
     2701 ctgtatgtgc gcagctcgcg ttgcattttc ggaagcgctc gttttcggaa acgctttgaa
     2761 gttcctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaaa
     2821 accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag
     2881 ctactaaaat attgcgaata ccgcttccac aaacattgct caaaagtatc tctttgctat
     2941 atatctctgt gctatatccc tatataacct acccatccac ctttcgctcc ttgaacttgc
     3001 atctaaactc gacctctaca ttttttatgt ttatctctag tattactctt tagacaaaaa
     3061 aattgtagta agaactattc atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc
     3121 ctatacgtag tatatagaga caaaatagaa gaaaccgttc ataattttct gaccaatgaa
     3181 gaatcatcaa cgctatcact ttctgttcac aaagtatgcg caatccacat cggtatagaa
     3241 tataatcggg gatgccttta tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta
     3301 aacgcgggaa gtggagtcag gcttttttta tggaagagaa aatagacacc aaagtagcct
     3361 tcttctaacc ttaacggacc tacagtgcaa aaagttatca agagactgca ttatagagcg
     3421 cacaaaggag aaaaaaagta atctaagatg ctttgttaga aaaatagcgc tctcgggatg
     3481 catttttgta gaacaaaaaa gaagtataga ttctttgttg gtaaaatagc gctctcgcgt
     3541 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
     3601 cgttgcattt ttgttttaca aaaatgaagc acagattctt cgttggtaaa atagcgcttt
     3661 cgcgttgcat ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc
     3721 tctcgcgttg catttttgtt ctacaaaatg aagcacagat gcttcgttga attcgtaatc
     3781 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3841 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3901 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3961 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4021 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4081 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4141 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4201 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4261 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4321 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4381 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4441 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4501 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4561 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4621 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4681 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4741 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4801 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4861 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4921 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4981 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5041 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5101 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5161 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5221 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5281 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5341 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5401 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5461 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5521 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5581 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5641 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5701 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5761 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5821 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5881 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5941 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6001 tcgtc
//
