LOCUS       sequence-435382-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI forTF12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7113)
  TITLE     Regulating transcription factors in Yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7113)
  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            "Unpublished"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: pCfb13202
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        459..533
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(938..971)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1178..1197)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1203..1222)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        1264..1355
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1356..1424,1425..2216)
                     /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(1574..1593)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2338..2926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2827..2846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3080..3097
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3195..3212
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3196..3212
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3682..3715
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             4857..5885
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             4857..5885
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     primer_bind     complement(5902..5920)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 ttcgtagata atggaataca aatggatatc cagagtatac acatggatag tatacactga
       61 cacgacaatt ctgtatctct ttatgttaac tactgtgagg cattaaatag agcttgatat
      121 ataaaatgtt acatttcaca gtctgaactt ttgcagatta cctaatttgg taagatatta
      181 attatgaact gaaagttgat ggcatcccta aatttgatgc aacggaatgc gtgcgatagt
      241 gaatcattgc taacagatca agtaatcgta tcaatcgttc cacgtcttat ccagactcga
      301 tttccgccct aacctaagga aactgctgag taatttcgag gttggttcct aacaaaagcc
      361 aagaaacgcg ccgatggttt agtggtaaaa tccatcgttg ccatcgatgg gcccccggtt
      421 cgattccggg tcggcgcatt ccaacctcca tcatcacagt tttagagcta gaaatagcaa
      481 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtggtgcttt
      541 ttttgtgtgg gtacggttga tttcgataga gcagcccatt gaaccttgat agtggcattt
      601 gaaatgagtg tagaaaaggc agtggtggac ttatagatcc aggaactaaa aatatacatc
      661 aatggtagtt agttttacag atttatctat gtagaaatgt gatatgacca gaatttcaag
      721 actgaagagc tcttcaatgt ccatcaactt ttcccacacc ctgtggtctc ccagtgacac
      781 ggtcaaagac gtggccgaga ctctgggcat ctccaatctg cccgatgacg tggccaaaac
      841 gcttgccatg gacgtggaat accgaatcca cgaggtgatt gaacaggcgc tcaagttcat
      901 cgcgtgcatt cgcggccgca tttaaatccc cacttcagaa gttcctatac tttctagaga
      961 ataggaactt cactatagag tcgaataagg gcgacacccc ataattagcc cgggcgaaag
     1021 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
     1081 atggggagtc cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg
     1141 gtcaggtggg accaccgcgc tactgccgcc aggcaaacaa ggggtgttat gagccatatt
     1201 caggtataaa tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc
     1261 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1321 gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat ttccgtgtcg
     1381 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1441 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1501 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1561 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1621 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1681 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1741 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1801 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1861 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcgatggca acaacgttgc
     1921 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1981 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2041 ttgctgataa atccggagcc ggtgagcgtg gttctcgcgg tatcatcgca gcgctggggc
     2101 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2161 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaagcag
     2221 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     2281 tacgcgcgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttagccc accacttcaa
     2521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2821 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2881 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2941 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3001 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3061 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaggcg agagtaggga
     3121 actgccaggc atcaaactaa gcagaaggcc cctgacggat ggcctttttg cgtttctaca
     3181 aactctttct gtgttgtaaa acgacggcca gtcttaagct cgggccccct gggcggttct
     3241 gataacgagt aatcgttaat ccgcaaataa cgtaaaaacc cgcttcggcg ggttttttta
     3301 tggggggagt ttagggaaag agcatttgtc agaatattta agggcgcctg tcactttgct
     3361 tgatatatga gaattattta accttataaa tgagaaaaaa gcaacgcact ttaaataaga
     3421 tacgttgctt tttcgattga tgaacaccta taattaaact attcatctat tatttatgat
     3481 tttttgtata tacaatattt ctagtttgtt aaagagaatt aagaaaataa atctcgaaaa
     3541 taataaaggg aaaatcagtt tttgatatca aaattataca tgtcaacgat aatacaaaat
     3601 ataatacaaa ctataagatg ttatcagtat ttattatgca tttagaataa attttgtgtc
     3661 gcccttattc gactcactat agaagttcct attctctaga aagtatagga acttcacttc
     3721 attttattta aatttgcggc cgctatgtct gataaaagga tgtaacatag gcaagctgct
     3781 cgtgagtgtt gagtacgaac cttagatcca aatcacccgc acccacggat atacttgctt
     3841 gaatatacag tagtaaggag tttggcgccc gttttttcga gccccacacg tttcggtgag
     3901 tatgagcggc ggcagattcg agcgtttccg gtttccgcgg ctggacgaga gcccatgatg
     3961 ggggctccca ccaccagcaa tcagggccct gattacacac ccacctgtaa tgtcatgctg
     4021 ttcatcgtgg ttaatgctgc tgtgtgctgt gtgtgtgtgt tgtttggcgc tcattgttgc
     4081 gttatgcagc gtacaccaca atattggaag cttattagcc tttctatttt ttcgtttgca
     4141 aggcttaaca acattgctgt ggagagggat ggggatatgg aggccgctgg agggagtcgg
     4201 agaggcgttt tggagcggct tggcctggcg cccagctcgc gaaacgcacc taggaccctt
     4261 tggcacgccg aaatgtgcca cttttcagtc tagtaacgcc ttacctacgt cattccatgc
     4321 atgcatgttt gcgccttttt tcccttgccc ttgatcgcca cacagtacag tgcactgtac
     4381 agtggaggtt ttgggggggt cttagatggg agctaaaagc ggcctagcgg tacactagtg
     4441 ggattgtatg gagtggcatg gagcctaggt ggagcctgac aggacgcacg accggctagc
     4501 ccgtgacaga cgatgggtgg ctcctgttgt ccaccgcgta caaatgtttg ggccaaagtc
     4561 ttgtcagcct tgcttgcgaa cctaattccc aattttgtca cttcgcaccc ccattgatcg
     4621 agccctaacc cctgcccatc aggcaatcca attaagctcg cattgtctgc cttgtttagt
     4681 ttggctcctg cccgtttcgg cgtccacttg cacaaacaca aacaagcatt atatataagg
     4741 ctcgtctctc cctcccaacc acactcactt ttttgcccgt cttcccttgc taacacaaaa
     4801 gtcaagaaca caaacaacca ccccaacccc cttacacaca agacatatct acagcaatgg
     4861 gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     4921 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     4981 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     5041 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     5101 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     5161 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gcaatggatg
     5221 ccattgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     5281 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     5341 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     5401 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     5461 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     5521 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     5581 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     5641 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     5701 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     5761 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     5821 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     5881 aataatcatg taattagtta tgtcacgctt acattcacgc cctcccccca catccgctct
     5941 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt tttttatagt
     6001 tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacagacg
     6061 cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg gacgctcgag
     6121 caggcttgga ggcgacgtgg cagtggctcg cgggaggatc ggcgggaaac tccgggatat
     6181 ccgtcgagag tttacacgtg aatgggcagc gcaatccgtt gacgacgata cgactggcaa
     6241 agtagcgacg atacctgcca gacaggtgac atgtgcaggc cgcactaaca aggaaacggg
     6301 cgctgggggg ggcgggcttc tagactttgc ccttgaacag gaatctagtg ggggcttgtc
     6361 tttccgccaa tgggggagcg cctgttgagc gaccgtgcat gctggaacgc caagtgtatg
     6421 tacagctggt ggtctcgcag cggtatgtga cgggacttac atctctcgtt ttttcatgac
     6481 cacgttttca caggctcgga ggtacgttaa agttttgaag gctgcatctg aaccgaggta
     6541 tgggggagtt tgaacagcaa cagtgttggg gctgaggggg ccaagatcgg ggcaagcaga
     6601 ggtcttagat caattgtggg gatcccaaag ggctcgttat cacctttttc cacccaattc
     6661 gggtcccaat tgatccacta ctggcttgcc caagttaccc cagaaatgcc gccccggatt
     6721 tctccaaaaa cctaataagc ttcatggaac ttggtggaag tgactttcta cagagtggag
     6781 agaaccgtgg acacgtggca atggcgctga ccgtgtcccc gagccgaatc gacgtgaggg
     6841 gagaacggag tatctgcggt catgtgacct tccagagcgg cgtcgccagt gtgcacgcgg
     6901 tgacccccag tttgggtctc tgtcacacgc atactacctc ggctctccac atgctgaact
     6961 ttatctttcg tggggatcat accgaaagtt gcaactacca ggtgtatata aagcctggta
     7021 gactcccccc actttggacc tcatccaacc aagacacaca aaaatggacc gacttaacaa
     7081 cctcgccacc cagctcgagc agaaccccgc caa
//
