LOCUS       sequence-320307-        8483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pmig-13::Zip(-)::QFAD::NLS::SL2::mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8483)
  AUTHORS   Mao S, Qi Y, Zhu H, Huang X, Zou Y, Chi T
  TITLE     A Tet/Q Hybrid System for Robust and Versatile Control of Transgene 
            Expression in C. elegans.
  JOURNAL   iScience. 2019 Jan 25;11:224-237. doi: 10.1016/j.isci.2018.12.023. 
            Epub 2018 Dec 27.
  PUBMED    30634168
REFERENCE   2  (bases 1 to 8483)
  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: "iScience.";
            date: "2019-01-25"; volume: "11:224-237. doi"; pages: " 
            10.1016/j.isci.2018.12.023. Epub 2018 Dec 27"
COMMENT     Sequence Label: TC512
FEATURES             Location/Qualifiers
     source          1..8483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      208..796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     697..716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     950..967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1084..1105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1120..1125,1126..1143,1144..1150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1158..1174
                     /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     1163..1185
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             5325..5345
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5605..6315
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5750..5768)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6018..6037
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7246..7268)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7368..7387
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7469..7487)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7555..7659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7660..7728,7729..8483,1..37)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7878..7897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
       61 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      121 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      181 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      241 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      301 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      361 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      421 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      481 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      541 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca
      601 ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      661 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      721 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      781 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      841 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      901 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      961 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1021 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1081 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1141 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctgggggtg
     1201 gggggggggt tgggggtggg gtgttgttgd gggtgtgtdd tttgtdatga ccatgattac
     1261 gccaagcttg catgcgcggc cggtaccctg acactaagtt tctctgcagt gttacacgaa
     1321 ctttttacaa tctactacta ctaacgaacg tcgcttgtat gtgttgcgct ctgaaaagtt
     1381 cacatctgtt gctgcctttt gtacattatt tgagaatctt tactcatttc gtgttctttt
     1441 atcaatttag tgaataaact gattcatatt gattaacaag tagttgaaca aaaagaaacg
     1501 gtgtcctttt gtgtttgcat tttatctctg tcctttgtag tctgatttta attttttctt
     1561 actctattca tctccttctc tgctccacca aaccggtata tatgactagg gcttacacag
     1621 acgtgcttca ttttcttttc ttgcttcttc cccacaaatg atacactgtc aatgggcgag
     1681 gaagaagagc tgcttcttgg gacggactcg tcgttctttc tcatcaatca ccggcgatgc
     1741 actcgacaca aacacgttac aagaaagacc ctcccgggct ctttcagccc attttgttga
     1801 aatatgtgct tttgaaacgc aacattcgaa tgatttgtca cgacagtcgg cctaattaat
     1861 tgactctcga aaatgagcat acgattgagt gttaatgtgc cgaagatgta gggaggagca
     1921 agaaaagagg gtggaagaga tgatgagtga cagactgtct taacagatga cctgcttgtg
     1981 gtggtcacct gtcgagttat gatctcagta attgcttcaa ttttccacat acgttcccaa
     2041 gtgtctcatt agaatataaa taagacaata ttttgttctg tgtcctgtag agaatacaca
     2101 attccaacta gaaacgtttt tttttcattt ttttaaattg atttcaaaac atcttttgac
     2161 ctgatgtcag tgtgggggat ttttttggca tttgtaatcg aaattaatga atttttagac
     2221 aatttctgaa aaatagtaag gtttaaaaaa atcttaagga cggcacacac tgtacaacaa
     2281 tactttgact gatggttttt ctgtttttca ttacaatctt attttttcta gtcagctttt
     2341 ttatgtgatt atttaaaaaa acaatggtct tgctttagat agtcaagatc tcacactctt
     2401 aataaaaaag gttttaatac ttcagatttt gctttaactt ttggaaattg ccaaccgttt
     2461 ctgaaaaatt tgaagtcttt gaaaacaatt acagcacttt tgcacgtttt gctgctatca
     2521 aatgtttaat ataaataagc aaaatatttg tgaaaaataa aatacttcac ttcatttttt
     2581 aaattttgct tctttctcac aaatgatata ttgactgtaa atgtcagaat tttgaaaagt
     2641 taaaaaaaat tatgttgaaa atttattttt ttttcttact gacagactga tgtgcctttt
     2701 tcagcatctt gaataagttc aagttttttt aatcgaagct gtttccagtg tctgtcagtg
     2761 acccacacac tagaacgcct aaaataaaaa tcgaaatttt tggagtatcc gaatttaatt
     2821 tggaatcaaa ttgatgcttg aaatatatgt gagctcttaa aataataata aaaaattgct
     2881 acaatcattt tattaaaata tttttcgatc taaaaattgt tcatcattga gtaatgtaat
     2941 tttgataaat tagacccccc ccccttcttc tacaaaactg atcaaaaggt ccgaaagact
     3001 tagaagaccc gcacaatagt agaatattca atggccgaat agtgaatgcg cgaggcgggt
     3061 ggttgctgtc ttctcggttc atataaggct catctgttca atcaccaaat gatcgatgtg
     3121 tttttgtggc cttcggcgtc agtttggagc agcacttcaa cctcattgaa acctctgtct
     3181 tcctcctatc tttgcctttt tttagtcaag aggcggtgtg aacgacgcaa tattcccgtt
     3241 ggttatcacc acactagaag tttttttgag cactcttatc agggaacaat tattgatgat
     3301 ttattgccaa ctagtgggtt ggttttgcta ttttttgaaa tttactggta tggtcttttg
     3361 agattttcag gcagttaaaa cttactaaga ctgatggtta acaaaaagtt gtaatataca
     3421 atactaactg aagcgaaact ttgaacgaaa aatatattaa actgttaaat tttatttgca
     3481 gctagctaac taataatttg tatgtttagt gcatttaaaa aagttttcag ctatattata
     3541 atgatagttt gaaactatga taaaactgag tttatacgac tttttgaata cacaaaaaaa
     3601 atttaaaaat tttttgaaaa cttttttaat gtattttttg tagtatttta ttttccctgg
     3661 cctaggaatt atgaaaaaaa gtttctagat ttcaatttat tccttatttt gacaaattat
     3721 tatgaaataa ttcattaatt tcattttgta cgtaaatggg cgggactaat gagatcctca
     3781 aactccttgt gattctcttt cctcctcgcc ttcttctacc atactcctgg acctcgctct
     3841 tcccgcgcac tctcttcccc ctctttcact cttgccacac ttgccaaggg agtcataaat
     3901 cgtgcaacat gtttagtagg tgccgttttg aaatatagat gctctttttt tctttctcac
     3961 aaaccatctt ttagttggtc tcgctcatct attttgaaaa atgacgatcc gtgtagagct
     4021 ttgattcttc atagttattt tcttgtgtgt gcgtcgggcc tcatactgcc aggtagagtg
     4081 tacgtgttta cctaacaagg gagaggtcta tgtggcactg cgcacttcat cgtatgcaaa
     4141 tttgctttta tgatctcttc cctcaggtca ttgtccatca taagcatcca cacacttttt
     4201 ttttggtgtt ttcttctctt tttttgggtc accagttcac aaccaaaaat ataggtggtc
     4261 aaaacataac atttttctgt acttacagta agtggaaatg tcgtctaagc acacttagaa
     4321 tattctaaat aaaagagaat tattagaaaa caactttaaa atgtttctgg caacgttttg
     4381 aaagtttgag aaatttataa caaacgttca aatcaagttg caacataatt agaatataaa
     4441 atttgtgaaa aacacttttt ggtcaacttc caaaattatc aatgatgaaa aagtgactgt
     4501 ttgcaactat ttggctttaa attaaagaac taatttttta ttttaatttt cactatttcc
     4561 ctacaagcaa cctgtaaagt gatattgtgt gtgactattt tttctctttt accacaatat
     4621 ttgagtatta accccccttt agctgtcttc aattatcatt taattcagaa tttcaggtaa
     4681 tggcttctga acaattggaa aagaaacttc aagctttgga aaagaaactt gcacaattgg
     4741 aatggaagaa ccaagctctt gaaaagaaat tggcacaagg atcaggatct ggatcaggat
     4801 ctggatcagg tacccttgag cttccgccga ctgcgactgc cactgcaagt atcatgcccc
     4861 atgtcatgga acagcccctg tcaacgtcaa taaatccggt caatgatcgg tttaatggaa
     4921 taccgaatcc taccccctac aatagcgatg cggccctaga cgcaattact caaaccaatg
     4981 actacggatc agtcaatacc catggcattc tcagtacata tccaccacct gccacccact
     5041 tgaacgaggc ctcggtggcc cttgcgcctg gaggtgctcc tcccagacca cccccgccat
     5101 acgttgatag tacaacgaac catcctcctt accatagtaa cctcgtcccc atggccaatt
     5161 tcggatattc gactgtagac tatgatgcga tggtggatga tctggcatcg atcgagtaca
     5221 cggatgcggt tgacgttgat ccgcagttca tgacgaacct cgggtttgtc ccagggtgta
     5281 attttagcga tatcaacacg tatgagcaac catggggcag tgggccaaag aagaagcgta
     5341 aggtctaagc tgtctcatcc tactttcacc tagttaactg cttgtcttaa aatctatgct
     5401 tctctttagt atctaaaatt ttcctagaag cttacaagta tataaatggt ctcttctcaa
     5461 taaaggttgt atatttattc atcttattga atctgccatt tcctcgtttt tgcgagttta
     5521 tataccttcc aattttcttt ctattgtatt ttcaacttct aattttaatt cagggaaact
     5581 gcttcaacgc atccggtaga aaaaatggtg agcaagggcg aggaggataa catggccatc
     5641 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     5701 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     5761 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     5821 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     5881 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     5941 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     6001 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     6061 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     6121 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     6181 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     6241 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     6301 gagctgtaca agtaacattc gtagaattcc aactgagcgc cggtcgctac cattaccaac
     6361 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     6421 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     6481 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     6541 ttgttattat caaaaaaact tcttcttaat ttctttgttt tttagcttct tttaagtcac
     6601 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     6661 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     6721 gtgtacactt cttatgtttt ttttacttct gataaatttt ttttgaaaca tcatagaaaa
     6781 aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc aatttttatt
     6841 tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa aagttttgga
     6901 gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct
     6961 ttttgggggt ttcccctatt gtttgtcaag agtttcgagg acggcgtttt tcttgctaaa
     7021 atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct
     7081 cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg
     7141 ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttccta ctagtcggcc
     7201 gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7261 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7321 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7381 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7441 ataccgcatc aggcggcctt aattaagggc ctcgtgatac gcctattttt ataggttaat
     7501 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     7561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7861 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     8041 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     8101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     8161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     8221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     8281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     8341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     8401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8461 atggatgaac gaaatagaca gat
//
