LOCUS       sequence-424054-        4497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GPD promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4497)
  AUTHORS   Jiang W, Wang S, Avila P, Jorgensen TS, Yang Z, Borodina I
  TITLE     Combinatorial iterative method for metabolic engineering of Yarrowia
            lipolytica: application for betanin biosynthesis.
  JOURNAL   Metab Eng. 2024 Sep 9:S1096-7176(24)00110-1. doi: 
            10.1016/j.ymben.2024.09.003.
  PUBMED    39260817
REFERENCE   2  (bases 1 to 4497)
  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: "Metab Eng. 
            2024 Sep 9:S1096-7176(24)00110-1. doi: 10.1016/j.ymben.2024.09.003."
COMMENT     Sequence Label: pCfB12708
FEATURES             Location/Qualifiers
     source          1..4497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(559..577)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(560..577)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(595..611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(595..611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(608..630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    619..635
                     /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)."
     promoter        complement(join(643..649,650..667,668..673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    688..709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(826..843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(997..1585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1077..1096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1723..2097)
                     /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"
     regulatory      2094..2103
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2303..3097)
                     /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             complement(2303..3097)
                     /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(2414..2433)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3024..3043
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(3446..3748)
                     /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     complement(3548..3567)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3871..3893
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3885..3902
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3886..3902
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3909..3928
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3909..3927
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctggttcgga cgcagtccgc cgcggcgtat cgatatccgc aaggttccat ggcgccattg
       61 ccctccgtcg gcgtctatcc cgcaacctct aaatagagcg ggaatataac ccaagcttct
      121 ttttttttcc tttaacacgc acacccccaa ctatcatgtt gctgctgctg tttgactcta
      181 ctctgtggag gggtgctccc acccaaccca acctacaggt ggatccggcg ctgtgattgg
      241 ctgataagtc tcctatccgg actaattctg accaatggga catgcgcgca ggacccaaat
      301 gccgcaatta cgtaacccca acgaaatgcc tacccctctt tggagcccag cggccccaaa
      361 tccccccaag cagcccggtt ctaccggctt ccatctccaa gcaccccttt ctccacaccc
      421 cacaaaaaga cccgtgcagg acatcctact gcgtcccgtt gagaccgatg agacgatgca
      481 agggcgaatt ccagcacact ggcggccgtt actagtggat ccgagctcgg taccaagctt
      541 gatgcatagc ttgagtattc tatagtgtca cctaaatagc ttggcgtaat catggtcata
      601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      661 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1681 cacctagatc cttttaaatt aaaaatgaag ttttagcacg tgtcagtcct gctcctcggc
     1741 cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
     1801 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
     1861 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
     1921 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
     1981 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
     2041 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt
     2101 ggccctcctc acgtgctatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2221 agtgccacct gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     2281 ggaaattgta agcgttaata attcagaaga actcgtcaag aaggcgatag aaggcgatgc
     2341 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     2401 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     2461 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     2521 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     2581 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     2641 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     2701 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     2761 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     2821 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     2881 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     2941 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     3001 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     3061 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     3121 cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     3181 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     3241 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     3301 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     3361 ggggtcagca ccgtttctgc ggactggctt tctacgtgaa aaggatctag gtgaagatcc
     3421 tttttgataa tctcatgcct gacatttata ttccccagaa catcaggtta atggcgtttt
     3481 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
     3541 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
     3601 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
     3661 gtcgccccgg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
     3721 ctcttttata ggtgtaaacc ttaaactgcc gtacgtatag gctgcgcaac tgttgggaag
     3781 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3841 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3901 gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca tgctcgagcg
     3961 gccgccagtg tgatggatat ctgcagaatt cgcccttatg ccgtctcagg tctcacattt
     4021 gttgatgtgt gtttaattca agaatgaata tagagaagag aagaagaaaa aagattcaat
     4081 tgagccggcg atgcagaccc ttatataaat gttgccttgg acagacggag caagcccgcc
     4141 caaacctacg ttcggtataa tatgttaagc tttttaacac aaaggtttgg cttggggtaa
     4201 cctgatgtgg tgcaaaagac cgggcgttgg cgagccattg cgcgggcgaa tggggccgtg
     4261 actcgcctca aattcgaggg cgtgcctcaa ttcgtgcccc cgtggctttt tcccgccgtt
     4321 tccgccccgt ttgcaccact gcagccgctt ctttggttcg gacaccttgc tgcgagctag
     4381 gtgccttgtg ctacttaaaa agtggcctcc caacaccaac atgacatgag tgcgtgggcc
     4441 aagacacgtt ggcggggtcg cagtcggctc aatggcccgg aaaaaacgct gctggag
//
