LOCUS       sequence-435638-        4496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrFBA with the overhangs for SapI restriction site can be insert 
            into the base plasmid of TUNEYALI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4496)
  TITLE     Regulating transcription factors in Yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4496)
  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            "Unpublished"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: pCfb12565
FEATURES             Location/Qualifiers
     source          1..4496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    234..255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(270..275,276..293,294..300)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    308..324
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     313..335
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     332..348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     332..348
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        366..384
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     366..383
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1384..1403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1385..1403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1410..1427)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1410..1426)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1419..1441)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1564..1866
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1745..1764
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2215..3009
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2215..3009
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2269..2288)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2879..2898
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3209..3218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3215..3589
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3727..4315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4216..4235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
       61 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      121 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      181 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      241 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      301 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
      361 aagctattta ggtgacacta tagaatactc aagctatgca tcaagcttgg taccgagctc
      421 ggatccacta gtaacggccg ccagtgtgct ggaattcgcc cttgctcttc tcattgtgtg
      481 atgtgtagtt tagatttcga atctgtgggg aaagaaagga aaaaagagac tggcaaccga
      541 ttgggagagc cactgtttat atatacccta gacaagcccc ccgcttgtaa gatgttggtc
      601 aatgtaaacc agtattaagg ttggcaagtg caggagaagc aaggtgtggg taccgagcaa
      661 tggaaatgtg cggaaggcaa aaaaatgagg ccacggccta ttgtcggggc tatatccagg
      721 gggcgattga agtacactaa catgacatgt gtccacagac cctcaatctg gcctgatgag
      781 ccaaatccat acgcgctttc gcagctctaa aggctataac aagtcacacc accctgctcg
      841 acctcagcgc cttcactttt tgctaagaca aactgtacac gctgttccag cgttttctgc
      901 ctgcacctgg tgggacattt ggtgcaacct aaagtgctcg gaacctctgt ggtgtccaga
      961 tcagcgcagc agttccgagg tagttttgag gcccttagat gatgcaatgg tgtcagtcgc
     1021 tggatcacga gtcttaatgg cagtattcgt tcttatttgt gccattgagc cccgttatcc
     1081 tcgtatcttc taccccccat cccatccctt tgttggtgca accctaccca tttattgttg
     1141 ggtgcagccc aaccgacgtg gagagcttgg cttggccata taaaaaggcc cccccctagt
     1201 ggcaatggca gaaagtcagc tgtgagttgt tgaatttgtc atctaggcgg cctggccgtc
     1261 ttctccgggg caattgttcc tctatagtac tgcgtacact gttgtctgaa gagcaagggc
     1321 gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggcccaat
     1381 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     1441 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1501 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctatacgta
     1561 cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     1621 cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt
     1681 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     1741 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     1801 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     1861 atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag
     1921 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     1981 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     2041 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     2101 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     2161 tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2221 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2281 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2341 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     2401 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2461 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2521 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2581 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2641 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2701 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     2761 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2821 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     2881 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     2941 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3001 ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     3061 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3181 aatgcttcaa taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc
     3241 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     3301 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     3361 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     3421 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     3481 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     3541 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctaaa
     3601 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3661 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3841 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4441 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
