LOCUS       sequence-445694-        4563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EXP promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4563)
  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 4563)
  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: pCfB12706
FEATURES             Location/Qualifiers
     source          1..4563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      5..593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     494..513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     747..764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    881..902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(917..922,923..940,941..947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    955..971
                     /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     960..982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     979..995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     979..995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2225..2244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2226..2244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2251..2268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2251..2267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2260..2282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2405..2707
                     /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     2586..2605
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             3056..3850
                     /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             3056..3850
                     /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(3110..3129)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3720..3739
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      4050..4059
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4056..4430
                     /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"
ORIGIN
        1 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
       61 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      841 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      901 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      961 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctatttaggt
     1021 gacactatag aatactcaag ctatgcatca agcttggtac cgagctcgga tccactagta
     1081 acggccgcca gtgtgctgga attcgccctt atgccgtctc aggtctcaca tttgctgtag
     1141 atatgtcttg tgtgtaaggg ggttggggtg gttgtttgtg ttcttgactt ttgtgttagc
     1201 aagggaagac gggcaaaaaa gtgagtgtgg ttgggaggga gaaacgagcc ttatatataa
     1261 tgcttgtttg tgtttgtgca agtggacgcc gaaacgggca ggagccaaac taaacaaggc
     1321 agacaatgcg agcttaattg gattgcctga tgggcagggg ttagggctcg atcaatgggg
     1381 gtgcgaagtg acaaaattgg gaattaggtt cgcaagcaag gctgacaaga ctttggccca
     1441 aacatttgta cgcggtggac aacaggagcc acccatcgtc tgtcacgggc tagccggtcg
     1501 tgcgtcctgt caggctccac ccaggctcca tgccactcca tacaatccca ctagtgtacc
     1561 gctaggccgc ttttagctcc catctaagac ccccccaaaa cctccactgt acagtgcact
     1621 gtactgtgtg gcgatcaagg gcaagggaaa aaaggcgcaa acatgcatgc atggaatgac
     1681 gtaggtaagg cgttactaga ctgaaaagtg gcacatttcg gcgtgccaaa gggtcctagg
     1741 tgcgtttcgc gagtgggcgc caggccaagc cgctccaaaa cgcctctccg actccctcca
     1801 gcggcctcca tatccccatc cctctccaca gcaatgttgt taagccttgc aaacgaaaaa
     1861 atagaaaggc taataagctt ccaatattgt ggtgtacgct gcataacgca acaatgagcg
     1921 ccaaacaaca cacacacaca gcacacagca gcattaacca cgatgaacag catgacatta
     1981 caggtgggtg tgtaatcagg gccctgattg ctggtggtgg gagcccccat catgggctct
     2041 cgtcccgccg cggaaaccgg aaacgctcga atctgccgcc gctcatactc accgaaacgt
     2101 gtggggctcg aaaaaacggg cgccaaactc cccgttgaga ccgatgagac gatgcaaggg
     2161 cgaattctgc agatatccat cacactggcg gccgctcgag catgcatcta gagggcccaa
     2221 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     2281 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2341 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt
     2401 acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     2461 acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg
     2521 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     2581 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     2641 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     2701 aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta
     2761 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     2821 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     2881 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     2941 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     3001 ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat
     3061 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3121 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3181 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3241 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3301 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3361 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3421 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3481 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3541 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3601 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3661 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3721 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3781 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3841 gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     3901 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3961 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4021 aaatgcttca ataatagcac gtgaggaggg ccaccatggc caagttgacc agtgccgttc
     4081 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     4141 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     4201 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     4261 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     4321 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     4381 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctaa
     4441 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4501 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4561 gat
//
