LOCUS       sequence-435506-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI for TF48.
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  JOURNAL   Unpublished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            "Unpublished"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: pCfb13238
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             780..1808
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             780..1808
                     /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(1825..1843)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     misc_RNA        3495..3569
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(3974..4007)
                     /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(4214..4233)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4239..4258)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4300..4391
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4392..4460,4461..5252)
                     /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(4610..4629)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5374..5962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5863..5882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6116..6133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6231..6248
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6232..6248
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6718..6751
                     /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 tgcgttatgc agcgtacacc acaatattgg aagcttatta gcctttctat tttttcgttt
       61 gcaaggctta acaacattgc tgtggagagg gatggggata tggaggccgc tggagggagt
      121 cggagaggcg ttttggagcg gcttggcctg gcgcccagct cgcgaaacgc acctaggacc
      181 ctttggcacg ccgaaatgtg ccacttttca gtctagtaac gccttaccta cgtcattcca
      241 tgcatgcatg tttgcgcctt ttttcccttg cccttgatcg ccacacagta cagtgcactg
      301 tacagtggag gttttggggg ggtcttagat gggagctaaa agcggcctag cggtacacta
      361 gtgggattgt atggagtggc atggagccta ggtggagcct gacaggacgc acgaccggct
      421 agcccgtgac agacgatggg tggctcctgt tgtccaccgc gtacaaatgt ttgggccaaa
      481 gtcttgtcag ccttgcttgc gaacctaatt cccaattttg tcacttcgca cccccattga
      541 tcgagcccta acccctgccc atcaggcaat ccaattaagc tcgcattgtc tgccttgttt
      601 agtttggctc ctgcccgttt cggcgtccac ttgcacaaac acaaacaagc attatatata
      661 aggctcgtct ctccctccca accacactca cttttttgcc cgtcttccct tgctaacaca
      721 aaagtcaaga acacaaacaa ccaccccaac ccccttacac acaagacata tctacagcaa
      781 tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
      841 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
      901 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
      961 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     1021 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     1081 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg
     1141 atgccattgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     1201 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     1261 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     1321 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     1381 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     1441 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     1501 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     1561 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     1621 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     1681 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     1741 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     1801 aggaataatc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     1861 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     1921 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     1981 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     2041 gagcaggctt ggaggcgacg tggcagtggc tcgcgggagg atcggcggga aactccggga
     2101 tatccgtcga gagtttacac gtgaatgggc agcgcaatcc gttgacgacg atacgactgg
     2161 caaagtagcg acgatacctg ccagacaggt gacatgtgca ggccgcacta acaaggaaac
     2221 gggcgctggg gggggcgggc ttctagactt tgcccttgaa caggaatcta gtgggggctt
     2281 gtctttccgc caatggggga gcgcctgttg agcgaccgtg catgctggaa cgccaagtgt
     2341 atgtacagct ggtggtctcg cagcggtatg tgacgggact tacatctctc gttttttcat
     2401 gaccacgttt tcacaggctc ggaggtacgt taaagttttg aaggctgcat ctgaaccgag
     2461 gtatggggga gtttgaacag caacagtgtt ggggctgagg gggccaagat cggggcaagc
     2521 agaggtctta gatcaattgt ggggatccca aagggctcgt tatcaccttt ttccacccaa
     2581 ttcgggtccc aattgatcca ctactggctt gcccaagtta ccccagaaat gccgccccgg
     2641 atttctccaa aaacctaata agcttcatgg aacttggtgg aagtgacttt ctacagagtg
     2701 gagagaaccg tggacacgtg gcaatggcgc tgaccgtgtc cccgagccga atcgacgtga
     2761 ggggagaacg gagtatctgc ggtcatgtga ccttccagag cggcgtcgcc agtgtgcacg
     2821 cggtgacccc cagtttgggt ctctgtcaca cgcatactac ctcggctctc cacatgctga
     2881 actttatctt tcgtggggat cataccgaaa gttgcaacta ccaggtgtat ataaagcctg
     2941 gtagactccc cccactttgg acctcatcca accaagacac acaaaaatgg accgacttaa
     3001 caacctcgcc acccagctcg agcagaaccc cgccaattcg tagataatgg aatacaaatg
     3061 gatatccaga gtatacacat ggatagtata cactgacacg acaattctgt atctctttat
     3121 gttaactact gtgaggcatt aaatagagct tgatatataa aatgttacat ttcacagtct
     3181 gaacttttgc agattaccta atttggtaag atattaatta tgaactgaaa gttgatggca
     3241 tccctaaatt tgatgcaacg gaatgcgtgc gatagtgaat cattgctaac agatcaagta
     3301 atcgtatcaa tcgttccacg tcttatccag actcgatttc cgccctaacc taaggaaact
     3361 gctgagtaat ttcgaggttg gttcctaaca aaagccaaga aacgcgccga tggtttagtg
     3421 gtaaaatcca tcgttgccat cgatgggccc ccggttcgat tccgggtcgg cgcaacacaa
     3481 ggggaaaaac acaagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     3541 caacttgaaa aagtggcacc gagtcggtgg tgcttttttt gtgtgggtac ggtgccaatt
     3601 aaatcttgat ggacacaaac acccggagcg gcggggaggg gaaattaaac agcatgcatt
     3661 gggcgctgct ggcaacttgc gctcgaattg gcctcactat ggcatttccg tctggagatt
     3721 ttagactttg catgcatatt taatatcccg agagtgactg aagagctctt caatgtacaa
     3781 caacacacgg gggagctact ccgacgacac ccagacgtac tatgacgggt tccggcacga
     3841 cgacggcaaa aacgaccctc aggcgggatc catttcgacc cacataccac acaaccccca
     3901 caatgacatg cccagctcca tcgccattcc agaaatcgcg tgcattcgcg gccgcattta
     3961 aatccccact tcagaagttc ctatactttc tagagaatag gaacttcact atagagtcga
     4021 ataagggcga caccccataa ttagcccggg cgaaaggccc agtctttcga ctgagccttt
     4081 cgttttattt gatgcctggc agttccctac tctcgcatgg ggagtcccca cactaccatc
     4141 ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact
     4201 gccgccaggc aaacaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa
     4261 tgttcagaat tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac
     4321 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4381 aaaggaagaa tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4501 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4561 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4621 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4681 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4801 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4921 acaccacgat gcctgtagcg atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4981 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5041 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatcc ggagccggtg
     5101 agcgtggttc tcgcggtatc atcgcagcgc tggggccaga tggtaagccc tcccgtatcg
     5161 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5221 agataggtgc ctcactgatt aagcattggt aagcagagca ttacgctgac ttgacgggac
     5281 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     5341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5521 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     5581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6001 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6061 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6121 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     6181 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     6241 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     6301 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     6361 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     6421 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     6481 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     6541 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     6601 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     6661 cagtatttat tatgcattta gaataaattt tgtgtcgccc ttattcgact cactatagaa
     6721 gttcctattc tctagaaagt ataggaactt cacttcattt tatttaaatt tgcggccgct
     6781 atgtctgata aaaggatgta acataggcaa gctgctcgtg agtgttgagt acgaacctta
     6841 gatccaaatc acccgcaccc acggatatac ttgcttgaat atacagtagt aaggagtttg
     6901 gcgcccgttt tttcgagccc cacacgtttc ggtgagtatg agcggcggca gattcgagcg
     6961 tttccggttt ccgcggctgg acgagagccc atgatggggg ctcccaccac cagcaatcag
     7021 ggccctgatt acacacccac ctgtaatgtc atgctgttca tcgtggttaa tgctgctgtg
     7081 tgctgtgtgt gtgtgttgtt tggcgctcat tgt
//
