LOCUS       sequence-329941-        8424 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 8424)
  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 8424)
  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..8424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5065..5085
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5354..6064
                     /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(5499..5517)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5767..5786
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6980..7002)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7102..7121
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7203..7221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7289..7393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7394..7462,7463..8254)
                     /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(7612..7631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcgcggc
     1021 cggtaccctg acactaagtt tctctgcagt gttacacgaa ctttttacaa tctactacta
     1081 ctaacgaacg tcgcttgtat gtgttgcgct ctgaaaagtt cacatctgtt gctgcctttt
     1141 gtacattatt tgagaatctt tactcatttc gtgttctttt atcaatttag tgaataaact
     1201 gattcatatt gattaacaag tagttgaaca aaaagaaacg gtgtcctttt gtgtttgcat
     1261 tttatctctg tcctttgtag tctgatttta attttttctt actctattca tctccttctc
     1321 tgctccacca aaccggtata tatgactagg gcttacacag acgtgcttca ttttcttttc
     1381 ttgcttcttc cccacaaatg atacactgtc aatgggcgag gaagaagagc tgcttcttgg
     1441 gacggactcg tcgttctttc tcatcaatca ccggcgatgc actcgacaca aacacgttac
     1501 aagaaagacc ctcccgggct ctttcagccc attttgttga aatatgtgct tttgaaacgc
     1561 aacattcgaa tgatttgtca cgacagtcgg cctaattaat tgactctcga aaatgagcat
     1621 acgattgagt gttaatgtgc cgaagatgta gggaggagca agaaaagagg gtggaagaga
     1681 tgatgagtga cagactgtct taacagatga cctgcttgtg gtggtcacct gtcgagttat
     1741 gatctcagta attgcttcaa ttttccacat acgttcccaa gtgtctcatt agaatataaa
     1801 taagacaata ttttgttctg tgtcctgtag agaatacaca attccaacta gaaacgtttt
     1861 tttttcattt ttttaaattg atttcaaaac atcttttgac ctgatgtcag tgtgggggat
     1921 ttttttggca tttgtaatcg aaattaatga atttttagac aatttctgaa aaatagtaag
     1981 gtttaaaaaa atcttaagga cggcacacac tgtacaacaa tactttgact gatggttttt
     2041 ctgtttttca ttacaatctt attttttcta gtcagctttt ttatgtgatt atttaaaaaa
     2101 acaatggtct tgctttagat agtcaagatc tcacactctt aataaaaaag gttttaatac
     2161 ttcagatttt gctttaactt ttggaaattg ccaaccgttt ctgaaaaatt tgaagtcttt
     2221 gaaaacaatt acagcacttt tgcacgtttt gctgctatca aatgtttaat ataaataagc
     2281 aaaatatttg tgaaaaataa aatacttcac ttcatttttt aaattttgct tctttctcac
     2341 aaatgatata ttgactgtaa atgtcagaat tttgaaaagt taaaaaaaat tatgttgaaa
     2401 atttattttt ttttcttact gacagactga tgtgcctttt tcagcatctt gaataagttc
     2461 aagttttttt aatcgaagct gtttccagtg tctgtcagtg acccacacac tagaacgcct
     2521 aaaataaaaa tcgaaatttt tggagtatcc gaatttaatt tggaatcaaa ttgatgcttg
     2581 aaatatatgt gagctcttaa aataataata aaaaattgct acaatcattt tattaaaata
     2641 tttttcgatc taaaaattgt tcatcattga gtaatgtaat tttgataaat tagacccccc
     2701 ccccttcttc tacaaaactg atcaaaaggt ccgaaagact tagaagaccc gcacaatagt
     2761 agaatattca atggccgaat agtgaatgcg cgaggcgggt ggttgctgtc ttctcggttc
     2821 atataaggct catctgttca atcaccaaat gatcgatgtg tttttgtggc cttcggcgtc
     2881 agtttggagc agcacttcaa cctcattgaa acctctgtct tcctcctatc tttgcctttt
     2941 tttagtcaag aggcggtgtg aacgacgcaa tattcccgtt ggttatcacc acactagaag
     3001 tttttttgag cactcttatc agggaacaat tattgatgat ttattgccaa ctagtgggtt
     3061 ggttttgcta ttttttgaaa tttactggta tggtcttttg agattttcag gcagttaaaa
     3121 cttactaaga ctgatggtta acaaaaagtt gtaatataca atactaactg aagcgaaact
     3181 ttgaacgaaa aatatattaa actgttaaat tttatttgca gctagctaac taataatttg
     3241 tatgtttagt gcatttaaaa aagttttcag ctatattata atgatagttt gaaactatga
     3301 taaaactgag tttatacgac tttttgaata cacaaaaaaa atttaaaaat tttttgaaaa
     3361 cttttttaat gtattttttg tagtatttta ttttccctgg cctaggaatt atgaaaaaaa
     3421 gtttctagat ttcaatttat tccttatttt gacaaattat tatgaaataa ttcattaatt
     3481 tcattttgta cgtaaatggg cgggactaat gagatcctca aactccttgt gattctcttt
     3541 cctcctcgcc ttcttctacc atactcctgg acctcgctct tcccgcgcac tctcttcccc
     3601 ctctttcact cttgccacac ttgccaaggg agtcataaat cgtgcaacat gtttagtagg
     3661 tgccgttttg aaatatagat gctctttttt tctttctcac aaaccatctt ttagttggtc
     3721 tcgctcatct attttgaaaa atgacgatcc gtgtagagct ttgattcttc atagttattt
     3781 tcttgtgtgt gcgtcgggcc tcatactgcc aggtagagtg tacgtgttta cctaacaagg
     3841 gagaggtcta tgtggcactg cgcacttcat cgtatgcaaa tttgctttta tgatctcttc
     3901 cctcaggtca ttgtccatca taagcatcca cacacttttt ttttggtgtt ttcttctctt
     3961 tttttgggtc accagttcac aaccaaaaat ataggtggtc aaaacataac atttttctgt
     4021 acttacagta agtggaaatg tcgtctaagc acacttagaa tattctaaat aaaagagaat
     4081 tattagaaaa caactttaaa atgtttctgg caacgttttg aaagtttgag aaatttataa
     4141 caaacgttca aatcaagttg caacataatt agaatataaa atttgtgaaa aacacttttt
     4201 ggtcaacttc caaaattatc aatgatgaaa aagtgactgt ttgcaactat ttggctttaa
     4261 attaaagaac taatttttta ttttaatttt cactatttcc ctacaagcaa cctgtaaagt
     4321 gatattgtgt gtgactattt tttctctttt accacaatat ttgagtatta accccccttt
     4381 agctgtcttc aattatcatt taattcagaa tttcaggtaa tggcttctga acaattggaa
     4441 aagaaacttc aagctttgga aaagaaactt gcacaattgg aatggaagaa ccaagctctt
     4501 gaaaagaaat tggcacaagg atcaggatct ggatcaggat ctggatcagg tacccttgag
     4561 cttccgccga ctgcgactgc cactgcaagt atcatgcccc atgtcatgga acagcccctg
     4621 tcaacgtcaa taaatccggt caatgatcgg tttaatggaa taccgaatcc taccccctac
     4681 aatagcgatg cggccctaga cgcaattact caaaccaatg actacggatc agtcaatacc
     4741 catggcattc tcagtacata tccaccacct gccacccact tgaacgaggc ctcggtggcc
     4801 cttgcgcctg gaggtgctcc tcccagacca cccccgccat acgttgatag tacaacgaac
     4861 catcctcctt accatagtaa cctcgtcccc atggccaatt tcggatattc gactgtagac
     4921 tatgatgcga tggtggatga tctggcatcg atcgagtaca cggatgcggt tgacgttgat
     4981 ccgcagttca tgacgaacct cgggtttgtc ccagggtgta attttagcga tatcaacacg
     5041 tatgagcaac catggggcag tgggccaaag aagaagcgta aggtctaagc tgtctcatcc
     5101 tactttcacc tagttaactg cttgtcttaa aatctatgct tctctttagt atctaaaatt
     5161 ttcctagaag cttacaagta tataaatggt ctcttctcaa taaaggttgt atatttattc
     5221 atcttattga atctgccatt tcctcgtttt tgcgagttta tataccttcc aattttcttt
     5281 ctattgtatt ttcaacttct aattttaatt cagggaaact gcttcaacgc atccggtaga
     5341 aaaaatgctc gagatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     5401 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     5461 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5521 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     5581 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     5641 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     5701 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     5761 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     5821 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     5881 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     5941 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     6001 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     6061 gtaagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     6121 ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat
     6181 ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca
     6241 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     6301 tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     6361 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     6421 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     6481 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     6541 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     6601 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     6661 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     6721 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     6781 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     6841 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     6901 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     6961 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     7021 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     7081 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     7141 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaattaa
     7201 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     7261 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7321 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7381 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7441 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7501 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7561 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7621 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7681 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7741 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7801 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7861 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7921 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7981 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8041 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8101 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8161 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8221 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8401 cgtagaaaag atcaaaggat cttc
//
