LOCUS       sequence-435635-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI for TF36.
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  JOURNAL   Unpublished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            "Unpublished"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: pCfb13226
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        142..216
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(621..654)
                     /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(861..880)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(886..905)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        947..1038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1039..1107,1108..1899)
                     /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(1257..1276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2021..2609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2510..2529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2763..2780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2878..2895
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2879..2895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3365..3398
                     /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             4540..5568
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             4540..5568
                     /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(5585..5603)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 ggaaactgct gagtaatttc gaggttggtt cctaacaaaa gccaagaaac gcgccgatgg
       61 tttagtggta aaatccatcg ttgccatcga tgggcccccg gttcgattcc gggtcggcgc
      121 aaacagaggc gaccagtcct ggttttagag ctagaaatag caagttaaaa taaggctagt
      181 ccgttatcaa cttgaaaaag tggcaccgag tcggtggtgc tttttttgtg tgggtacggt
      241 caccacaagc gcccaccccg cctggtctgc cagtttatcc aaactaggca gtcacagagt
      301 cgagcgaacg gaacaccacc tcgccttcga ctcccccagc taccgctcgt taggttttga
      361 gtactcagag taagactttc ttaactccct tcacttcaaa ctgactgaag agctcttcaa
      421 tgatccgaac gttccactct accgagcgac gttcagccat tgtggtcaac aacaacaaca
      481 aggtgtccaa gccctcgccc aacttctcgt ccaagaagtt gtcgcctgtg gcccaggctc
      541 ctattgtcgt gtctgacgcg tcagactcct cggctattcg aatcgcgtgc attcgcggcc
      601 gcatttaaat ccccacttca gaagttccta tactttctag agaataggaa cttcactata
      661 gagtcgaata agggcgacac cccataatta gcccgggcga aaggcccagt ctttcgactg
      721 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gtccccacac
      781 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
      841 cgctactgcc gccaggcaaa caaggggtgt tatgagccat attcaggtat aaatgggctc
      901 gcgataatgt tcagaattgg ttaattggtt gtaacactga cccctatttg tttatttttc
      961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1021 tattgaaaaa ggaagaatat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1141 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1201 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1261 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1321 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1441 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1561 gagcgtgaca ccacgatgcc tgtagcgatg gcaacaacgt tgcgcaaact attaactggc
     1621 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatccgga
     1741 gccggtgagc gtggttctcg cggtatcatc gcagcgctgg ggccagatgg taagccctcc
     1801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1861 atcgctgaga taggtgcctc actgattaag cattggtaag cagagcatta cgctgacttg
     1921 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgc gcgtcgttcc
     1981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2161 atactgttct tctagtgtag ccgtagttag cccaccactt caagaactct gtagcaccgc
     2221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2581 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2641 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2701 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2761 gcagcgagtc agtgagcgag gaagcggaag gcgagagtag ggaactgcca ggcatcaaac
     2821 taagcagaag gcccctgacg gatggccttt ttgcgtttct acaaactctt tctgtgttgt
     2881 aaaacgacgg ccagtcttaa gctcgggccc cctgggcggt tctgataacg agtaatcgtt
     2941 aatccgcaaa taacgtaaaa acccgcttcg gcgggttttt ttatgggggg agtttaggga
     3001 aagagcattt gtcagaatat ttaagggcgc ctgtcacttt gcttgatata tgagaattat
     3061 ttaaccttat aaatgagaaa aaagcaacgc actttaaata agatacgttg ctttttcgat
     3121 tgatgaacac ctataattaa actattcatc tattatttat gattttttgt atatacaata
     3181 tttctagttt gttaaagaga attaagaaaa taaatctcga aaataataaa gggaaaatca
     3241 gtttttgata tcaaaattat acatgtcaac gataatacaa aatataatac aaactataag
     3301 atgttatcag tatttattat gcatttagaa taaattttgt gtcgccctta ttcgactcac
     3361 tatagaagtt cctattctct agaaagtata ggaacttcac ttcattttat ttaaatttgc
     3421 ggccgctatg tctgataaaa ggatgtaaca taggcaagct gctcgtgagt gttgagtacg
     3481 aaccttagat ccaaatcacc cgcacccacg gatatacttg cttgaatata cagtagtaag
     3541 gagtttggcg cccgtttttt cgagccccac acgtttcggt gagtatgagc ggcggcagat
     3601 tcgagcgttt ccggtttccg cggctggacg agagcccatg atgggggctc ccaccaccag
     3661 caatcagggc cctgattaca cacccacctg taatgtcatg ctgttcatcg tggttaatgc
     3721 tgctgtgtgc tgtgtgtgtg tgttgtttgg cgctcattgt tgcgttatgc agcgtacacc
     3781 acaatattgg aagcttatta gcctttctat tttttcgttt gcaaggctta acaacattgc
     3841 tgtggagagg gatggggata tggaggccgc tggagggagt cggagaggcg ttttggagcg
     3901 gcttggcctg gcgcccagct cgcgaaacgc acctaggacc ctttggcacg ccgaaatgtg
     3961 ccacttttca gtctagtaac gccttaccta cgtcattcca tgcatgcatg tttgcgcctt
     4021 ttttcccttg cccttgatcg ccacacagta cagtgcactg tacagtggag gttttggggg
     4081 ggtcttagat gggagctaaa agcggcctag cggtacacta gtgggattgt atggagtggc
     4141 atggagccta ggtggagcct gacaggacgc acgaccggct agcccgtgac agacgatggg
     4201 tggctcctgt tgtccaccgc gtacaaatgt ttgggccaaa gtcttgtcag ccttgcttgc
     4261 gaacctaatt cccaattttg tcacttcgca cccccattga tcgagcccta acccctgccc
     4321 atcaggcaat ccaattaagc tcgcattgtc tgccttgttt agtttggctc ctgcccgttt
     4381 cggcgtccac ttgcacaaac acaaacaagc attatatata aggctcgtct ctccctccca
     4441 accacactca cttttttgcc cgtcttccct tgctaacaca aaagtcaaga acacaaacaa
     4501 ccaccccaac ccccttacac acaagacata tctacagcaa tgggtaaaaa gcctgaactc
     4561 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     4621 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     4681 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     4741 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     4801 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     4861 gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg atgccattgc tgcggccgat
     4921 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     4981 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     5041 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     5101 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     5161 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     5221 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     5281 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     5341 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     5401 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     5461 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     5521 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaataatc atgtaattag
     5581 ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag
     5641 ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa
     5701 cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca
     5761 ttatactgaa aaccttgctt gagaaggttt tgggacgctc gagcaggctt ggaggcgacg
     5821 tggcagtggc tcgcgggagg atcggcggga aactccggga tatccgtcga gagtttacac
     5881 gtgaatgggc agcgcaatcc gttgacgacg atacgactgg caaagtagcg acgatacctg
     5941 ccagacaggt gacatgtgca ggccgcacta acaaggaaac gggcgctggg gggggcgggc
     6001 ttctagactt tgcccttgaa caggaatcta gtgggggctt gtctttccgc caatggggga
     6061 gcgcctgttg agcgaccgtg catgctggaa cgccaagtgt atgtacagct ggtggtctcg
     6121 cagcggtatg tgacgggact tacatctctc gttttttcat gaccacgttt tcacaggctc
     6181 ggaggtacgt taaagttttg aaggctgcat ctgaaccgag gtatggggga gtttgaacag
     6241 caacagtgtt ggggctgagg gggccaagat cggggcaagc agaggtctta gatcaattgt
     6301 ggggatccca aagggctcgt tatcaccttt ttccacccaa ttcgggtccc aattgatcca
     6361 ctactggctt gcccaagtta ccccagaaat gccgccccgg atttctccaa aaacctaata
     6421 agcttcatgg aacttggtgg aagtgacttt ctacagagtg gagagaaccg tggacacgtg
     6481 gcaatggcgc tgaccgtgtc cccgagccga atcgacgtga ggggagaacg gagtatctgc
     6541 ggtcatgtga ccttccagag cggcgtcgcc agtgtgcacg cggtgacccc cagtttgggt
     6601 ctctgtcaca cgcatactac ctcggctctc cacatgctga actttatctt tcgtggggat
     6661 cataccgaaa gttgcaacta ccaggtgtat ataaagcctg gtagactccc cccactttgg
     6721 acctcatcca accaagacac acaaaaatgg accgacttaa caacctcgcc acccagctcg
     6781 agcagaaccc cgccaattcg tagataatgg aatacaaatg gatatccaga gtatacacat
     6841 ggatagtata cactgacacg acaattctgt atctctttat gttaactact gtgaggcatt
     6901 aaatagagct tgatatataa aatgttacat ttcacagtct gaacttttgc agattaccta
     6961 atttggtaag atattaatta tgaactgaaa gttgatggca tccctaaatt tgatgcaacg
     7021 gaatgcgtgc gatagtgaat cattgctaac agatcaagta atcgtatcaa tcgttccacg
     7081 tcttatccag actcgatttc cgccctaacc taa
//
