LOCUS       sequence-435430-        7137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI for TF38.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  TITLE     Regulating transcription factors in Yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7137)
  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            "Unpublished"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: pCfb13228
FEATURES             Location/Qualifiers
     source          1..7137
                     /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(962..995)
                     /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(1202..1221)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1227..1246)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        1288..1379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1380..1448,1449..2240)
                     /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(1598..1617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2362..2950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2851..2870
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3104..3121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3219..3236
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3220..3236
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3706..3739
                     /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             4881..5909
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             4881..5909
                     /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(5926..5944)
                     /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 tcggcgcagg tgtgacctat gacccaaggt tttagagcta gaaatagcaa
      481 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtggtgcttt
      541 ttttgtgtgg gtacggtagc catcaacgag ttgtacaata ggaccaggtg agtacgagac
      601 cggagcgcgt cgacgcagac gatgacgaga cgcacacaga tgcagaatcc ccgacaccgg
      661 cgaacggccc caagccggcc ttcttgtaat gttttttgct tcccgtgaca cgacaagatg
      721 actgaagagc tcttcaatga gcttccccca acaagtaata gcgccgggcc aacggctcaa
      781 cgagcttctg gaggccatca aacaggagtt cgactccgtg accaacgagg cgtccgtcta
      841 ccggctgcac aaggacgagt ttgacgtcaa ggtgaaccag cagacgtcag atctgggcat
      901 cgcgtgcatt cgcggccgca atcgcgtgca ttcgcggccg caaatttaaa tccccacttc
      961 agaagttcct atactttcta gagaatagga acttcactat agagtcgaat aagggcgaca
     1021 ccccataatt agcccgggcg aaaggcccag tctttcgact gagcctttcg ttttatttga
     1081 tgcctggcag ttccctactc tcgcatgggg agtccccaca ctaccatcgg cgctacggcg
     1141 tttcacttct gagttcggca tggggtcagg tgggaccacc gcgctactgc cgccaggcaa
     1201 acaaggggtg ttatgagcca tattcaggta taaatgggct cgcgataatg ttcagaattg
     1261 gttaattggt tgtaacactg acccctattt gtttattttt ctaaatacat tcaaatatgt
     1321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaata
     1381 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1441 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1501 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1561 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1621 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1681 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1741 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1801 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1861 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1921 ctgtagcgat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1981 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2041 cggcccttcc ggctggctgg tttattgctg ataaatccgg agccggtgag cgtggttctc
     2101 gcggtatcat cgcagcgctg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2161 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2221 cactgattaa gcattggtaa gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2281 tgaccaaaat cccttaacgt gagttacgcg cgcgtcgttc cactgagcgt cagaccccgt
     2341 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2401 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2461 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2521 gccgtagtta gcccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2581 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2641 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2701 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2761 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2821 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2881 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2941 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3001 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3061 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3121 ggaagcggaa ggcgagagta gggaactgcc aggcatcaaa ctaagcagaa ggcccctgac
     3181 ggatggcctt tttgcgtttc tacaaactct ttctgtgttg taaaacgacg gccagtctta
     3241 agctcgggcc ccctgggcgg ttctgataac gagtaatcgt taatccgcaa ataacgtaaa
     3301 aacccgcttc ggcgggtttt tttatggggg gagtttaggg aaagagcatt tgtcagaata
     3361 tttaagggcg cctgtcactt tgcttgatat atgagaatta tttaacctta taaatgagaa
     3421 aaaagcaacg cactttaaat aagatacgtt gctttttcga ttgatgaaca cctataatta
     3481 aactattcat ctattattta tgattttttg tatatacaat atttctagtt tgttaaagag
     3541 aattaagaaa ataaatctcg aaaataataa agggaaaatc agtttttgat atcaaaatta
     3601 tacatgtcaa cgataataca aaatataata caaactataa gatgttatca gtatttatta
     3661 tgcatttaga ataaattttg tgtcgccctt attcgactca ctatagaagt tcctattctc
     3721 tagaaagtat aggaacttca cttcatttta tttaaattcg cggccgctat gtctgataaa
     3781 aggatgtaac ataggcaagc tgctcgtgag tgttgagtac gaaccttaga tccaaatcac
     3841 ccgcacccac ggatatactt gcttgaatat acagtagtaa ggagtttggc gcccgttttt
     3901 tcgagcccca cacgtttcgg tgagtatgag cggcggcaga ttcgagcgtt tccggtttcc
     3961 gcggctggac gagagcccat gatgggggct cccaccacca gcaatcaggg ccctgattac
     4021 acacccacct gtaatgtcat gctgttcatc gtggttaatg ctgctgtgtg ctgtgtgtgt
     4081 gtgttgtttg gcgctcattg ttgcgttatg cagcgtacac cacaatattg gaagcttatt
     4141 agcctttcta ttttttcgtt tgcaaggctt aacaacattg ctgtggagag ggatggggat
     4201 atggaggccg ctggagggag tcggagaggc gttttggagc ggcttggcct ggcgcccagc
     4261 tcgcgaaacg cacctaggac cctttggcac gccgaaatgt gccacttttc agtctagtaa
     4321 cgccttacct acgtcattcc atgcatgcat gtttgcgcct tttttccctt gcccttgatc
     4381 gccacacagt acagtgcact gtacagtgga ggttttgggg gggtcttaga tgggagctaa
     4441 aagcggccta gcggtacact agtgggattg tatggagtgg catggagcct aggtggagcc
     4501 tgacaggacg cacgaccggc tagcccgtga cagacgatgg gtggctcctg ttgtccaccg
     4561 cgtacaaatg tttgggccaa agtcttgtca gccttgcttg cgaacctaat tcccaatttt
     4621 gtcacttcgc acccccattg atcgagccct aacccctgcc catcaggcaa tccaattaag
     4681 ctcgcattgt ctgccttgtt tagtttggct cctgcccgtt tcggcgtcca cttgcacaaa
     4741 cacaaacaag cattatatat aaggctcgtc tctccctccc aaccacactc acttttttgc
     4801 ccgtcttccc ttgctaacac aaaagtcaag aacacaaaca accaccccaa cccccttaca
     4861 cacaagacat atctacagca atgggtaaaa agcctgaact caccgcgacg tctgtcgaga
     4921 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     4981 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     5041 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     5101 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     5161 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     5221 agccggtcgc ggaggcaatg gatgccattg ctgcggccga tcttagccag acgagcgggt
     5281 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     5341 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     5401 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     5461 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     5521 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     5581 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     5641 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     5701 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     5761 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     5821 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     5881 ccagcactcg tccgagggca aaggaataat catgtaatta gttatgtcac gcttacattc
     5941 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     6001 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     6061 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     6121 tgagaaggtt ttgggacgct cgagcaggct tggaggcgac gtggcagtgg ctcgcgggag
     6181 gatcggcggg aaactccggg atatccgtcg agagtttaca cgtgaatggg cagcgcaatc
     6241 cgttgacgac gatacgactg gcaaagtagc gacgatacct gccagacagg tgacatgtgc
     6301 aggccgcact aacaaggaaa cgggcgctgg ggggggcggg cttctagact ttgcccttga
     6361 acaggaatct agtgggggct tgtctttccg ccaatggggg agcgcctgtt gagcgaccgt
     6421 gcatgctgga acgccaagtg tatgtacagc tggtggtctc gcagcggtat gtgacgggac
     6481 ttacatctct cgttttttca tgaccacgtt ttcacaggct cggaggtacg ttaaagtttt
     6541 gaaggctgca tctgaaccga ggtatggggg agtttgaaca gcaacagtgt tggggctgag
     6601 ggggccaaga tcggggcaag cagaggtctt agatcaattg tggggatccc aaagggctcg
     6661 ttatcacctt tttccaccca attcgggtcc caattgatcc actactggct tgcccaagtt
     6721 accccagaaa tgccgccccg gatttctcca aaaacctaat aagcttcatg gaacttggtg
     6781 gaagtgactt tctacagagt ggagagaacc gtggacacgt ggcaatggcg ctgaccgtgt
     6841 ccccgagccg aatcgacgtg aggggagaac ggagtatctg cggtcatgtg accttccaga
     6901 gcggcgtcgc cagtgtgcac gcggtgaccc ccagtttggg tctctgtcac acgcatacta
     6961 cctcggctct ccacatgctg aactttatct ttcgtgggga tcataccgaa agttgcaact
     7021 accaggtgta tataaagcct ggtagactcc ccccactttg gacctcatcc aaccaagaca
     7081 cacaaaaatg gaccgactta acaacctcgc cacccagctc gagcagaacc ccgccaa
//
