LOCUS       sequence-435636-        7113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The base plasmid of TUNEYALI forTF07.
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  JOURNAL   Unpublished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            "Unpublished"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: pCfb13197
FEATURES             Location/Qualifiers
     source          1..7113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(740..773)
                     /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(1243..1260)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1243..1259)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1358..1375)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1529..2117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1609..1628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2239..3030,3031..3099))
                     /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     2862..2881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3100..3191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3233..3252
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3258..3277
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    3484..3517
                     /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)"
     misc_RNA        complement(3922..3996)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     5648..5666
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(5683..6711)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(5683..6711)
                     /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"
ORIGIN
        1 actccataca atcccactag tgtaccgcta ggccgctttt agctcccatc taagaccccc
       61 ccaaaacctc cactgtacag tgcactgtac tgtgtggcga tcaagggcaa gggaaaaaag
      121 gcgcaaacat gcatgcatgg aatgacgtag gtaaggcgtt actagactga aaagtggcac
      181 atttcggcgt gccaaagggt cctaggtgcg tttcgcgagc tgggcgccag gccaagccgc
      241 tccaaaacgc ctctccgact ccctccagcg gcctccatat ccccatccct ctccacagca
      301 atgttgttaa gccttgcaaa cgaaaaaata gaaaggctaa taagcttcca atattgtggt
      361 gtacgctgca taacgcaaca atgagcgcca aacaacacac acacacagca cacagcagca
      421 ttaaccacga tgaacagcat gacattacag gtgggtgtgt aatcagggcc ctgattgctg
      481 gtggtgggag cccccatcat gggctctcgt ccagccgcgg aaaccggaaa cgctcgaatc
      541 tgccgccgct catactcacc gaaacgtgtg gggctcgaaa aaacgggcgc caaactcctt
      601 actactgtat attcaagcaa gtatatccgt gggtgcgggt gatttggatc taaggttcgt
      661 actcaacact cacgagcagc ttgcctatgt tacatccttt tatcagacat agcggccgca
      721 aatttaaata aaatgaagtg aagttcctat actttctaga gaataggaac ttctatagtg
      781 agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt
      841 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
      901 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
      961 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     1021 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     1081 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     1141 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     1201 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     1261 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     1321 tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     1381 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1441 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1501 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1561 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1621 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1681 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1741 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1801 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1861 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1921 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     1981 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2041 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2101 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2161 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     2221 gtcagcgtaa tgctctgctt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2281 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2341 gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct caccggctcc
     2401 ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2461 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2521 agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc
     2581 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2641 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2701 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2761 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2821 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2881 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2941 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3001 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3061 aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt ttcaatatta
     3121 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3181 aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag
     3241 cccatttata cctgaatatg gctcataaca ccccttgttt gcctggcggc agtagcgcgg
     3301 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     3361 tggggactcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     3421 tcgaaagact gggcctttcg cccgggctaa ttatggggtg tcgcccttat tcgactctat
     3481 agtgaagttc ctattctcta gaaagtatag gaacttctga agtggggatt taaatgcggc
     3541 cgcgaatgca cgcgatcggc tgcaccgcca tatccatcat cggagttccc ggaccctgcc
     3601 tgttgccaaa gctaggcgat ggcgtagcca gatcgctctg cgcgtccatc tggcccatgc
     3661 ccgagtccat cattccgtcc atgaatccct ggggacccat gcccatgtcg ctttgcattg
     3721 aagagctctt cagtctttgc ttttttttag agttaggagc tgtggcgagc gcatgacgac
     3781 gagaaaacgc accctacgat ggggaccggg ttgcctgttg gggtggcgcc agtgtccctg
     3841 tccttgtgtc tgtgtggtcg tctcccgcga gtatgtatat gagtccagct cgctgttacc
     3901 gtacccacac aaaaaaagca ccaccgactc ggtgccactt tttcaagttg ataacggact
     3961 agccttattt taacttgcta tttctagctc taaaaccgtc taacacagtg aactcatgcg
     4021 ccgacccgga atcgaaccgg gggcccatcg atggcaacga tggattttac cactaaacca
     4081 tcggcgcgtt tcttggcttt tgttaggaac caacctcgaa attactcagc agtttcctta
     4141 ggttagggcg gaaatcgagt ctggataaga cgtggaacga ttgatacgat tacttgatct
     4201 gttagcaatg attcactatc gcacgcattc cgttgcatca aatttaggga tgccatcaac
     4261 tttcagttca taattaatat cttaccaaat taggtaatct gcaaaagttc agactgtgaa
     4321 atgtaacatt ttatatatca agctctattt aatgcctcac agtagttaac ataaagagat
     4381 acagaattgt cgtgtcagtg tatactatcc atgtgtatac tctggatatc catttgtatt
     4441 ccattatcta cgaattggcg gggttctgct cgagctgggt ggcgaggttg ttaagtcggt
     4501 ccatttttgt gtgtcttggt tggatgaggt ccaaagtggg gggagtctac caggctttat
     4561 atacacctgg tagttgcaac tttcggtatg atccccacga aagataaagt tcagcatgtg
     4621 gagagccgag gtagtatgcg tgtgacagag acccaaactg ggggtcaccg cgtgcacact
     4681 ggcgacgccg ctctggaagg tcacatgacc gcagatactc cgttctcccc tcacgtcgat
     4741 tcggctcggg gacacggtca gcgccattgc cacgtgtcca cggttctctc cactctgtag
     4801 aaagtcactt ccaccaagtt ccatgaagct tattaggttt ttggagaaat ccggggcggc
     4861 atttctgggg taacttgggc aagccagtag tggatcaatt gggacccgaa ttgggtggaa
     4921 aaaggtgata acgagccctt tgggatcccc acaattgatc taagacctct gcttgccccg
     4981 atcttggccc cctcagcccc aacactgttg ctgttcaaac tcccccatac ctcggttcag
     5041 atgcagcctt caaaacttta acgtacctcc gagcctgtga aaacgtggtc atgaaaaaac
     5101 gagagatgta agtcccgtca cataccgctg cgagaccacc agctgtacat acacttggcg
     5161 ttccagcatg cacggtcgct caacaggcgc tcccccattg gcggaaagac aagcccccac
     5221 tagattcctg ttcaagggca aagtctagaa gcccgccccc cccagcgccc gtttccttgt
     5281 tagtgcggcc tgcacatgtc acctgtctgg caggtatcgt cgctactttg ccagtcgtat
     5341 cgtcgtcaac ggattgcgct gcccattcac gtgtaaactc tcgacggata tcccggagtt
     5401 tcccgccgat cctcccgcga gccactgcca cgtcgcctcc aagcctgctc gagcgtccca
     5461 aaaccttctc aagcaaggtt ttcagtataa tgttacatgc gtacacgcgt ctgtacagaa
     5521 aaaaaagaaa aatttgaaat ataaataacg ttcttaatac taacataact ataaaaaaat
     5581 aaatagggac ctagacttca ggttgtctaa ctccttcctt ttcggttaga gcggatgtgg
     5641 ggggagggcg tgaatgtaag cgtgacataa ctaattacat gattattcct ttgccctcgg
     5701 acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca
     5761 gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac
     5821 gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct
     5881 ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccgcg gcgatcctgc
     5941 aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg
     6001 cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca
     6061 gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc
     6121 gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc
     6181 ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg
     6241 atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc
     6301 gaatgggccg aacccgctcg tctggctaag atcggccgca gcaatggcat ccattgcctc
     6361 cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc
     6421 ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aattccccaa tgtcaagcac
     6481 ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa
     6541 accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa
     6601 agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc
     6661 gatcagaaac ttctcgacag acgtcgcggt gagttcaggc tttttaccca ttgctgtaga
     6721 tatgtcttgt gtgtaagggg gttggggtgg ttgtttgtgt tcttgacttt tgtgttagca
     6781 agggaagacg ggcaaaaaag tgagtgtggt tgggagggag agacgagcct tatatataat
     6841 gcttgtttgt gtttgtgcaa gtggacgccg aaacgggcag gagccaaact aaacaaggca
     6901 gacaatgcga gcttaattgg attgcctgat gggcaggggt tagggctcga tcaatggggg
     6961 tgcgaagtga caaaattggg aattaggttc gcaagcaagg ctgacaagac tttggcccaa
     7021 acatttgtac gcggtggaca acaggagcca cccatcgtct gtcacgggct agccggtcgt
     7081 gcgtcctgtc aggctccacc taggctccat gcc
//
