LOCUS       sequence-435498-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI for TF26.
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  JOURNAL   Unpublished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            "Unpublished"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: pCfb13216
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             445..1473
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             445..1473
                     /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(1490..1508)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     misc_RNA        3160..3234
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(3639..3672)
                     /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(3879..3898)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3904..3923)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3965..4056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4057..4125,4126..4917)
                     /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(4275..4294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5039..5627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5528..5547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5781..5798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5896..5913
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5897..5913
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6383..6416
                     /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)"
ORIGIN
        1 ctaaaagcgg cctagcggta cactagtggg attgtatgga gtggcatgga gcctaggtgg
       61 agcctgacag gacgcacgac cggctagccc gtgacagacg atgggtggct cctgttgtcc
      121 accgcgtaca aatgtttggg ccaaagtctt gtcagccttg cttgcgaacc taattcccaa
      181 ttttgtcact tcgcaccccc attgatcgag ccctaacccc tgcccatcag gcaatccaat
      241 taagctcgca ttgtctgcct tgtttagttt ggctcctgcc cgtttcggcg tccacttgca
      301 caaacacaaa caagcattat atataaggct cgtctctccc tcccaaccac actcactttt
      361 ttgcccgtct tcccttgcta acacaaaagt caagaacaca aacaaccacc ccaaccccct
      421 tacacacaag acatatctac agcaatgggt aaaaagcctg aactcaccgc gacgtctgtc
      481 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
      541 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
      601 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
      661 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
      721 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
      781 ctgcagccgg tcgcggaggc aatggatgcc attgctgcgg ccgatcttag ccagacgagc
      841 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
      901 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
      961 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     1021 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     1081 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     1141 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     1201 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     1261 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     1321 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     1381 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     1441 cgccccagca ctcgtccgag ggcaaaggaa taatcatgta attagttatg tcacgcttac
     1501 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     1561 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     1621 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     1681 tgcttgagaa ggttttggga cgctcgagca ggcttggagg cgacgtggca gtggctcgcg
     1741 ggaggatcgg cgggaaactc cgggatatcc gtcgagagtt tacacgtgaa tgggcagcgc
     1801 aatccgttga cgacgatacg actggcaaag tagcgacgat acctgccaga caggtgacat
     1861 gtgcaggccg cactaacaag gaaacgggcg ctgggggggg cgggcttcta gactttgccc
     1921 ttgaacagga atctagtggg ggcttgtctt tccgccaatg ggggagcgcc tgttgagcga
     1981 ccgtgcatgc tggaacgcca agtgtatgta cagctggtgg tctcgcagcg gtatgtgacg
     2041 ggacttacat ctctcgtttt ttcatgacca cgttttcaca ggctcggagg tacgttaaag
     2101 ttttgaaggc tgcatctgaa ccgaggtatg ggggagtttg aacagcaaca gtgttggggc
     2161 tgagggggcc aagatcgggg caagcagagg tcttagatca attgtgggga tcccaaaggg
     2221 ctcgttatca cctttttcca cccaattcgg gtcccaattg atccactact ggcttgccca
     2281 agttacccca gaaatgccgc cccggatttc tccaaaaacc taataagctt catggaactt
     2341 ggtggaagtg actttctaca gagtggagag aaccgtggac acgtggcaat ggcgctgacc
     2401 gtgtccccga gccgaatcga cgtgagggga gaacggagta tctgcggtca tgtgaccttc
     2461 cagagcggcg tcgccagtgt gcacgcggtg acccccagtt tgggtctctg tcacacgcat
     2521 actacctcgg ctctccacat gctgaacttt atctttcgtg gggatcatac cgaaagttgc
     2581 aactaccagg tgtatataaa gcctggtaga ctccccccac tttggacctc atccaaccaa
     2641 gacacacaaa aatggaccga cttaacaacc tcgccaccca gctcgagcag aaccccgcca
     2701 attcgtagat aatggaatac aaatggatat ccagagtata cacatggata gtatacactg
     2761 acacgacaat tctgtatctc tttatgttaa ctactgtgag gcattaaata gagcttgata
     2821 tataaaatgt tacatttcac agtctgaact tttgcagatt acctaatttg gtaagatatt
     2881 aattatgaac tgaaagttga tggcatccct aaatttgatg caacggaatg cgtgcgatag
     2941 tgaatcattg ctaacagatc aagtaatcgt atcaatcgtt ccacgtctta tccagactcg
     3001 atttccgccc taacctaagg aaactgctga gtaatttcga ggttggttcc taacaaaagc
     3061 caagaaacgc gccgatggtt tagtggtaaa atccatcgtt gccatcgatg ggcccccggt
     3121 tcgattccgg gtcggcgcat ttgcacactc aagacatggg ttttagagct agaaatagca
     3181 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtggtgctt
     3241 tttttgtgtg ggtacggtat actggaacga ttatctatcg attcatctac tttttactac
     3301 cacttttcat acaaaaatgc gtgtcgatca ggtgtcaaga gcaagcaacg aaatgtcact
     3361 gcctctattt tttgggcatt ttttcgcgtg tctcattttt tgccctaccc acgttgtcgg
     3421 gactgaagag ctcttcaatg actgaccgac ttttcttggc caacctcaat gcgattgagg
     3481 atcctcaaca taccgagcct tcgcccgaca acgatgatct atccatcttc acgaacacgc
     3541 agttcttcga ctttgacatg ggctgctcca ccgacattgc tgcttctgtg gacgacatga
     3601 tcgcgtgcat tcgcggccgc atttaaatcc ccacttcaga agttcctata ctttctagag
     3661 aataggaact tcactataga gtcgaataag ggcgacaccc cataattagc ccgggcgaaa
     3721 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     3781 catggggagt ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg
     3841 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaaca aggggtgtta tgagccatat
     3901 tcaggtataa atgggctcgc gataatgttc agaattggtt aattggttgt aacactgacc
     3961 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4021 tgataaatgc ttcaataata ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc
     4081 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4141 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4201 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4261 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4321 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4381 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4441 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4501 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4561 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcgatggc aacaacgttg
     4621 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4681 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4741 attgctgata aatccggagc cggtgagcgt ggttctcgcg gtatcatcgc agcgctgggg
     4801 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4861 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaagca
     4921 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     4981 ttacgcgcgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5161 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttagcc caccacttca
     5221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaggc gagagtaggg
     5821 aactgccagg catcaaacta agcagaaggc ccctgacgga tggccttttt gcgtttctac
     5881 aaactctttc tgtgttgtaa aacgacggcc agtcttaagc tcgggccccc tgggcggttc
     5941 tgataacgag taatcgttaa tccgcaaata acgtaaaaac ccgcttcggc gggttttttt
     6001 atggggggag tttagggaaa gagcatttgt cagaatattt aagggcgcct gtcactttgc
     6061 ttgatatatg agaattattt aaccttataa atgagaaaaa agcaacgcac tttaaataag
     6121 atacgttgct ttttcgattg atgaacacct ataattaaac tattcatcta ttatttatga
     6181 ttttttgtat atacaatatt tctagtttgt taaagagaat taagaaaata aatctcgaaa
     6241 ataataaagg gaaaatcagt ttttgatatc aaaattatac atgtcaacga taatacaaaa
     6301 tataatacaa actataagat gttatcagta tttattatgc atttagaata aattttgtgt
     6361 cgcccttatt cgactcacta tagaagttcc tattctctag aaagtatagg aacttcactt
     6421 cattttattt aaatttgcgg ccgctatgtc tgataaaagg atgtaacata ggcaagctgc
     6481 tcgtgagtgt tgagtacgaa ccttagatcc aaatcacccg cacccacgga tatacttgct
     6541 tgaatataca gtagtaagga gtttggcgcc cgttttttcg agccccacac gtttcggtga
     6601 gtatgagcgg cggcagattc gagcgtttcc ggtttccgcg gctggacgag agcccatgat
     6661 gggggctccc accaccagca atcagggccc tgattacaca cccacctgta atgtcatgct
     6721 gttcatcgtg gttaatgctg ctgtgtgctg tgtgtgtgtg ttgtttggcg ctcattgttg
     6781 cgttatgcag cgtacaccac aatattggaa gcttattagc ctttctattt tttcgtttgc
     6841 aaggcttaac aacattgctg tggagaggga tggggatatg gaggccgctg gagggagtcg
     6901 gagaggcgtt ttggagcggc ttggcctggc gcccagctcg cgaaacgcac ctaggaccct
     6961 ttggcacgcc gaaatgtgcc acttttcagt ctagtaacgc cttacctacg tcattccatg
     7021 catgcatgtt tgcgcctttt ttcccttgcc cttgatcgcc acacagtaca gtgcactgta
     7081 cagtggaggt tttggggggg tcttagatgg gag
//
