LOCUS       sequence-437825-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI for TF30.
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  JOURNAL   Unpublished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            "Unpublished"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: pCfb13220
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             469..1497
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             469..1497
                     /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(1514..1532)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     misc_RNA        3184..3258
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(3663..3696)
                     /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(3903..3922)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3928..3947)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3989..4080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4081..4149,4150..4941)
                     /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(4299..4318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5063..5651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5552..5571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5805..5822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5920..5937
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5921..5937
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6407..6440
                     /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 ttttgggggg gtcttagatg ggagctaaaa gcggcctagc ggtacactag tgggattgta
       61 tggagtggca tggagcctag gtggagcctg acaggacgca cgaccggcta gcccgtgaca
      121 gacgatgggt ggctcctgtt gtccaccgcg tacaaatgtt tgggccaaag tcttgtcagc
      181 cttgcttgcg aacctaattc ccaattttgt cacttcgcac ccccattgat cgagccctaa
      241 cccctgccca tcaggcaatc caattaagct cgcattgtct gccttgttta gtttggctcc
      301 tgcccgtttc ggcgtccact tgcacaaaca caaacaagca ttatatataa ggctcgtctc
      361 tccctcccaa ccacactcac ttttttgccc gtcttccctt gctaacacaa aagtcaagaa
      421 cacaaacaac caccccaacc cccttacaca caagacatat ctacagcaat gggtaaaaag
      481 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
      541 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
      601 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
      661 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
      721 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
      781 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga tgccattgct
      841 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
      901 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
      961 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     1021 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     1081 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     1141 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     1201 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     1261 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     1321 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     1381 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     1441 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaataatca
     1501 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     1561 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     1621 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     1681 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg agcaggcttg
     1741 gaggcgacgt ggcagtggct cgcgggagga tcggcgggaa actccgggat atccgtcgag
     1801 agtttacacg tgaatgggca gcgcaatccg ttgacgacga tacgactggc aaagtagcga
     1861 cgatacctgc cagacaggtg acatgtgcag gccgcactaa caaggaaacg ggcgctgggg
     1921 ggggcgggct tctagacttt gcccttgaac aggaatctag tgggggcttg tctttccgcc
     1981 aatgggggag cgcctgttga gcgaccgtgc atgctggaac gccaagtgta tgtacagctg
     2041 gtggtctcgc agcggtatgt gacgggactt acatctctcg ttttttcatg accacgtttt
     2101 cacaggctcg gaggtacgtt aaagttttga aggctgcatc tgaaccgagg tatgggggag
     2161 tttgaacagc aacagtgttg gggctgaggg ggccaagatc ggggcaagca gaggtcttag
     2221 atcaattgtg gggatcccaa agggctcgtt atcacctttt tccacccaat tcgggtccca
     2281 attgatccac tactggcttg cccaagttac cccagaaatg ccgccccgga tttctccaaa
     2341 aacctaataa gcttcatgga acttggtgga agtgactttc tacagagtgg agagaaccgt
     2401 ggacacgtgg caatggcgct gaccgtgtcc ccgagccgaa tcgacgtgag gggagaacgg
     2461 agtatctgcg gtcatgtgac cttccagagc ggcgtcgcca gtgtgcacgc ggtgaccccc
     2521 agtttgggtc tctgtcacac gcatactacc tcggctctcc acatgctgaa ctttatcttt
     2581 cgtggggatc ataccgaaag ttgcaactac caggtgtata taaagcctgg tagactcccc
     2641 ccactttgga cctcatccaa ccaagacaca caaaaatgga ccgacttaac aacctcgcca
     2701 cccagctcga gcagaacccc gccaattcgt agataatgga atacaaatgg atatccagag
     2761 tatacacatg gatagtatac actgacacga caattctgta tctctttatg ttaactactg
     2821 tgaggcatta aatagagctt gatatataaa atgttacatt tcacagtctg aacttttgca
     2881 gattacctaa tttggtaaga tattaattat gaactgaaag ttgatggcat ccctaaattt
     2941 gatgcaacgg aatgcgtgcg atagtgaatc attgctaaca gatcaagtaa tcgtatcaat
     3001 cgttccacgt cttatccaga ctcgatttcc gccctaacct aaggaaactg ctgagtaatt
     3061 tcgaggttgg ttcctaacaa aagccaagaa acgcgccgat ggtttagtgg taaaatccat
     3121 cgttgccatc gatgggcccc cggttcgatt ccgggtcggc gcagcgtcag caggaaagtc
     3181 gaggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     3241 agtggcaccg agtcggtggt gctttttttg tgtgggtacg gtctcgcata gcacaactca
     3301 cccaactcat gcaccagttt ttttcatcaa cgtcgcataa atccgatgtg gggtagggtc
     3361 cactcgatga atgatataga cgaggtggtc gatactggta ttgtggcgta ttatataaac
     3421 atcgagaacc gctgccgaac tccagactga agagctcttc aatggtgaag cgaaaatttg
     3481 aagacgagga ccagaccttg cgagagggcc agagcggctt tgctactgtg agtgactacg
     3541 agagcggccc accaagccat tgacgccgtt atcccttggt tatctctttg gctaacacag
     3601 atgaacaaca actcgtactg gaaatcgcgt gcattcgcgg ccgcatttaa atccccactt
     3661 cagaagttcc tatactttct agagaatagg aacttcacta tagagtcgaa taagggcgac
     3721 accccataat tagcccgggc gaaaggccca gtctttcgac tgagcctttc gttttatttg
     3781 atgcctggca gttccctact ctcgcatggg gagtccccac actaccatcg gcgctacggc
     3841 gtttcacttc tgagttcggc atggggtcag gtgggaccac cgcgctactg ccgccaggca
     3901 aacaaggggt gttatgagcc atattcaggt ataaatgggc tcgcgataat gttcagaatt
     3961 ggttaattgg ttgtaacact gacccctatt tgtttatttt tctaaataca ttcaaatatg
     4021 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat
     4081 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4141 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4201 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4261 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4321 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4381 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4441 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4501 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4561 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4621 cctgtagcga tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4681 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4741 tcggcccttc cggctggctg gtttattgct gataaatccg gagccggtga gcgtggttct
     4801 cgcggtatca tcgcagcgct ggggccagat ggtaagccct cccgtatcgt agttatctac
     4861 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4921 tcactgatta agcattggta agcagagcat tacgctgact tgacgggacg gcgcaagctc
     4981 atgaccaaaa tcccttaacg tgagttacgc gcgcgtcgtt ccactgagcg tcagaccccg
     5041 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5101 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5161 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5221 agccgtagtt agcccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5281 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5341 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5401 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5461 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5521 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5581 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5641 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5701 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5761 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5821 aggaagcgga aggcgagagt agggaactgc caggcatcaa actaagcaga aggcccctga
     5881 cggatggcct ttttgcgttt ctacaaactc tttctgtgtt gtaaaacgac ggccagtctt
     5941 aagctcgggc cccctgggcg gttctgataa cgagtaatcg ttaatccgca aataacgtaa
     6001 aaacccgctt cggcgggttt ttttatgggg ggagtttagg gaaagagcat ttgtcagaat
     6061 atttaagggc gcctgtcact ttgcttgata tatgagaatt atttaacctt ataaatgaga
     6121 aaaaagcaac gcactttaaa taagatacgt tgctttttcg attgatgaac acctataatt
     6181 aaactattca tctattattt atgatttttt gtatatacaa tatttctagt ttgttaaaga
     6241 gaattaagaa aataaatctc gaaaataata aagggaaaat cagtttttga tatcaaaatt
     6301 atacatgtca acgataatac aaaatataat acaaactata agatgttatc agtatttatt
     6361 atgcatttag aataaatttt gtgtcgccct tattcgactc actatagaag ttcctattct
     6421 ctagaaagta taggaacttc acttcatttt atttaaattt gcggccgcta tgtctgataa
     6481 aaggatgtaa cataggcaag ctgctcgtga gtgttgagta cgaaccttag atccaaatca
     6541 cccgcaccca cggatatact tgcttgaata tacagtagta aggagtttgg cgcccgtttt
     6601 ttcgagcccc acacgtttcg gtgagtatga gcggcggcag attcgagcgt ttccggtttc
     6661 cgcggctgga cgagagccca tgatgggggc tcccaccacc agcaatcagg gccctgatta
     6721 cacacccacc tgtaatgtca tgctgttcat cgtggttaat gctgctgtgt gctgtgtgtg
     6781 tgtgttgttt ggcgctcatt gttgcgttat gcagcgtaca ccacaatatt ggaagcttat
     6841 tagcctttct attttttcgt ttgcaaggct taacaacatt gctgtggaga gggatgggga
     6901 tatggaggcc gctggaggga gtcggagagg cgttttggag cggcttggcc tggcgcccag
     6961 ctcgcgaaac gcacctagga ccctttggca cgccgaaatg tgccactttt cagtctagta
     7021 acgccttacc tacgtcattc catgcatgca tgtttgcgcc ttttttccct tgcccttgat
     7081 cgccacacag tacagtgcac tgtacagtgg agg
//
