LOCUS       sequence-248790-        4724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for PCR amplification of XdCrtYB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4724)
  AUTHORS   Milne N, Tramontin LRR, Borodina I
  TITLE     A teaching protocol demonstrating the use of EasyClone and 
            CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae 
            and Yarrowia lipolytica.
  JOURNAL   FEMS Yeast Res. 2019 Sep 26. pii: foz062. doi: 
            10.1093/femsyr/foz062.
  PUBMED    31556952
REFERENCE   2  (bases 1 to 4724)
  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: "FEMS Yeast 
            Res. 2019 Sep 26. pii: foz062. doi: 10.1093/femsyr/foz062."
COMMENT     Sequence Label: pCfB8739
FEATURES             Location/Qualifiers
     source          1..4724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2324..2341)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2324..2340)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2333..2355)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2549..2568)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2668..2690
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2728..2746)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2814..2918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2919..2987,2988..3779)
                     /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     complement(3137..3156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3950..4538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4439..4458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4692..4709
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc
      241 ctgcaggtcg actctagagg atccccatct gtcatatgac ggctctcgca tattaccaga
      301 tccatctgat ctatactctc ccaattcttg gtcttctcgg cctgctcact tccccgattt
      361 tgacaaaatt tgacatctac aaaatatcga tcctcgtatt tattgcgttt agtgcaacca
      421 caccatggga ctcatggatc atcagaaatg gcgcatggac atatccatca gcggagagtg
      481 gccaaggcgt gtttggaacg tttctagatg ttccatatga agagtacgct ttctttgtca
      541 ttcaaaccgt aatcaccggc ttggtctacg tcttggcaac taggcacctt ctcccatctc
      601 tcgcgcttcc caagactaga tcgtccgccc tttctctcgc gctcaaggcg ctcatccctc
      661 tgcccattat ctacctattt accgctcacc ccagcccatc gcccgacccg ctcgtgacag
      721 atcactactt ctacatgcgg gcactctcct tactcatcac cccacctacc atgctcttgg
      781 cagcattatc aggcgaatat gctttcgatt ggaaaagtgg ccgagcaaag tcaactattg
      841 caacaatcat gatcccgacg gtgtatctga tttgggtaga ttatgttgct gtcggtcaag
      901 actcttggtc gatcaacgat gagaagattg tagggtggag gcttggaggt gtactaccca
      961 ttgaggaagc tatgttcttc ttactgacga atctaatgat tgttctgggt ctgtctgcct
     1021 gcgatcatac tcaggcccta tacctgctac acggtcgaac tatttatggc aacaaaaaga
     1081 tgccatcttc atttcccctc attacaccgc ctgtgctctc cctgtttttt agcagccgac
     1141 catactcttc tcagccaaaa cgtgacttgg aactggcagt caagttgttg gagaaaaaga
     1201 gccggagctt ttttgttgcc tcggctggat ttcctagcga agttagggag aggctggttg
     1261 gactatacgc attctgccgg gtgactgatg atcttatcga ctctcctgaa gtatcttcca
     1321 acccgcatgc cacaattgac atggtctccg attttcttac cctactattt gggcccccgc
     1381 tacacccttc gcaacctgac aagatccttt cttcgccttt acttcctcct tcgcaccctt
     1441 cccgacccac gggaatgtat cccctcccgc ctcctccttc gctctcgcct gccgagctcg
     1501 ttcaattcct taccgaaagg gttcccgttc aataccattt cgccttcagg ttgctcgcta
     1561 agttgcaagg gctgatccct cgatacccac tcgacgaact ccttagagga tacaccactg
     1621 atcttatctt tcccttatcg acagaggcag tccaggctcg gaagacgcct atcgagacca
     1681 cagctgactt gctggactat ggtctatgtg tagcaggctc agtcgccgag ctattggtct
     1741 atgtctcttg ggcaagtgca ccaagtcagg tccctgccac catagaagaa agagaagctg
     1801 tgttagtggc aagccgagag atgggaactg cccttcagtt ggtgaacatt gctagggaca
     1861 ttaaagggga cgcaacagaa gggagatttt acctaccact ctcattcttt ggtcttcggg
     1921 atgaatcaaa gcttgcgatc ccgactgatt ggacggaacc tcggcctcaa gatttcgaca
     1981 aactcctcag tctatctcct tcgtccacat taccatcttc aaacgcctca gaaagcttcc
     2041 ggttcgaatg gaagacgtac tcgcttccat tagtcgccta cgcagaggat cttgccaaac
     2101 attcttataa gggaattgac cgacttccta ccgaggttca agcgggaatg cgagcggctt
     2161 gcgcgagcta cctactgatc ggccgagaga tcaaagtcgt ttggaaagga gacgtcggag
     2221 agagaaggac agttgccgga tggaggagag tacggaaagt cttgagtgtg gtcatgagcg
     2281 gatgggaagg gcagtaaatc gcgtgggtac cgagctcgaa ttcactggcc gtcgttttac
     2341 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2401 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2461 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2521 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2581 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2641 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2701 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2761 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2821 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2881 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2941 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3001 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3061 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3121 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3181 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3241 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3301 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3361 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3421 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3481 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3541 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3601 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3661 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3721 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3781 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3841 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3901 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3961 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4021 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4081 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4141 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4201 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4261 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4321 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4381 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4501 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4561 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4621 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4681 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgc
//
