LOCUS       sequence-435639-        4604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PrPGM 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 4604)
  TITLE     Regulating transcription factors in Yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4604)
  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            "Unpublished"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: pCfb12561
FEATURES             Location/Qualifiers
     source          1..4604
                     /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(1492..1511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1493..1511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1518..1535)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1518..1534)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1527..1549)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1672..1974
                     /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     1853..1872
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2323..3117
                     /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             2323..3117
                     /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(2377..2396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2987..3006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3317..3326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3323..3697
                     /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      3835..4423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4324..4343
                     /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 agaccagtat
      481 gtacagtagc gacagtatct tccatacttc actttacagt aaattaaaga atacaccaaa
      541 actcgttctc aagctcttgt caaaaaactc caaaaaaata taaaatatat aatatattat
      601 atatatcgcg atacctcatt aattctcacg tgacacagat tattaacgtc tcgtaccaac
      661 cacagattac gacccattcg cagtcacagt tcactagggt ttgggttgca tccgttgaga
      721 gtggtttgtt tttaaccttc tccatgtgct cactcaggtt ttgggttcag atcaaatcaa
      781 ggcgtgaacc actgtttgag gacaaatgtg acacaaccaa ccagtgtcag gggcaagtcc
      841 gtgacaaagg ggaagataca atgcaattac tgacagttac ggactgcctc gatgccctaa
      901 ccttgcccca aaataagaca actgtcctcg tttaagcgca accctattca gcgtcacgtc
      961 ataatagcgt ttggatagca ctagtctatg aggagcgttt tatgttgcgg tgagggcgat
     1021 tggtgctcat atgggttcaa ttgaggtggt ggaacgagct tagtcttcaa ttgaggtgcg
     1081 agcgacacaa ttgggtgtca cgtggcctaa ttgacctcgg atcgtggagt ccccagttat
     1141 acagcaacca cgaggtgcat gagtaggaga cgtcaccaga caatagggtt tttttggact
     1201 ggagagggta gggcaaaagc gctcaacggg ctgtttgggg agctatgggg gaggaattgg
     1261 cgatatttgt gaggttgacg gctccgattt gcgtgttttg tcgcttctgc atctccccat
     1321 acccatatct tccctcccca cctctttcca cgataatttt acggatcagc aataaggttc
     1381 cttctcctag tttccacgtc catatatatc tatgagaaga gcaagggcga attctgcaga
     1441 tatccatcac actggcggcc gctcgagcat gcatctagag ggcccaattc gccctatagt
     1501 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1561 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1621 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tatacgtacg gcagtttaag
     1681 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     1741 attgacacgc cggggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     1801 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     1861 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     1921 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     1981 ggcatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa agccagtccg
     2041 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     2101 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     2161 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     2221 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     2281 ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2341 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2401 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2461 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     2521 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2581 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2641 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2701 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     2761 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     2821 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     2881 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     2941 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3001 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3061 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     3121 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3181 caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3241 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3301 atagcacgtg aggagggcca ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     3361 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     3421 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     3481 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     3541 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     3601 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     3661 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctaaaac ttcattttta
     3721 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3781 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3841 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3901 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3961 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4021 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4081 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4141 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4201 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4261 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4321 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4381 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4441 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4501 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4561 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
