LOCUS       sequence-487419-        4519 bp    DNA     circular SYN 29-SEP-2025
DEFINITION  PrTPI 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 4519)
  TITLE     Regulating transcription factors in Yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4519)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCfb12620
FEATURES             Location/Qualifiers
     source          1..4519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      141..150
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             147..521
                     /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      659..1247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1148..1167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1401..1418
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1535..1556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1571..1576,1577..1594,1595..1601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1609..1625
                     /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     1614..1636
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1633..1649
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1633..1649
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1667..1685
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1667..1684
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2835..2854)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2836..2854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2861..2878)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2861..2877)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2870..2892)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3015..3317
                     /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     3196..3215
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3666..4460
                     /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             3666..4460
                     /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(3720..3739)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4330..4349
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
       61 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      121 aataatagca cgtgaggagg gccaccatgg ccaagttgac cagtgccgtt ccggtgctca
      181 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
      241 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
      301 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
      361 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
      421 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
      481 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta aaacttcatt
      541 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      601 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      661 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      721 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      781 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      841 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      901 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      961 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1021 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1081 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1141 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1201 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1261 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1321 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1381 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     1441 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1501 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     1561 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     1621 aacaatttca cacaggaaac agctatgacc atgattacgc caagctattt aggtgacact
     1681 atagaatact caagctatgc atcaagcttg gtaccgagct cggatccact agtaacggcc
     1741 gccagtgtgc tggaattcgc ccttgctctt cagacttagt tgactggaaa agtcacctgg
     1801 gggttcattt ctggtgttac aagaatggaa gaacattgag atgtagttta gtagatggag
     1861 aagacttgag ttctaaacaa aagagctgaa atcatatcct tcagtagtag tatagtcctg
     1921 ttatcacagc atcaattacc cccgtccaag taagttgatt gggatttttg tttacagata
     1981 cagtaatata cttgactatt tctttacagg tgactcagaa agtgcatgtt ggaaatgagc
     2041 cacagaccaa gacaagatat gacaaaattg cactattcga tgcagaattc gacggtgttt
     2101 ccattggtgt tatgacattc atctgcattc atacaaaaaa gtcttggtag tggtactttt
     2161 gcgttattac ctccgatatc tacgcacccc ccaacccccc tgctacagta aagagtgtga
     2221 gtctactgta catgcttact aaaccaccta ctgtacagcg aaacccctca gcaaaatcac
     2281 acaatcagct cattacaaca cacccaatga cctcaccaca aattctatac gccttttgac
     2341 gccattatta cagtagcttg caacgccgtt gtcttaggtt ccatttttag tgctctatta
     2401 cctcacttaa cccgtatagg cagatcaggc catggcacta agtgtagagc tagaggttga
     2461 tatcgccacg agtgctccat cagggctagg gtggggttag aaatacagtc cgtgcgcact
     2521 caaaaggcgt ccgggttagg gcatccgata atatcgcctg gactcggcgc catattctcg
     2581 acttctgggc gcgttgtatt catctcctcc gcttcccaac acttccaccc gtttctccat
     2641 cccaaccaat agaatagggt aaccttattc gggacacttt cgtcatacat agtcagatat
     2701 acaagcaatg tcactctcct tcgtactcgt acatacaaca caactacatt caaaatgaga
     2761 agagcaaggg cgaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     2821 gagggcccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     2881 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2941 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     3001 gcctatacgt acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     3061 ttgtggatgt acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg
     3121 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     3181 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
     3241 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     3301 tgttctgggg aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc
     3361 ttttcacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     3421 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     3481 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     3541 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     3601 cgccaaggat ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt
     3661 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3721 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3781 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3841 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3901 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3961 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4021 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4081 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4141 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     4201 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4261 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4321 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4381 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4441 ttcttgacga gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc
     4501 ttacgcatct gtgcggtat
//
