LOCUS       sequence-329944-        9283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pdpy30::loxP::mcherry::let-858terminator::loxP::GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9283)
  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 9283)
  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: TC646
FEATURES             Location/Qualifiers
     source          1..9283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1074..1090
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1074..1090
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(2371..2404)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3702..3735)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3797..3825)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4573..4593
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      4637..4960
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(5073..5090)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5073..5089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5911..6705
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5965..5984)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6575..6594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7406..7422
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7407..7423
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8078..8100
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8138..8156)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8224..8328
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8329..8397,8398..9189)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8547..8566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggt cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgcaccgtac gtctcgagga attctatcag gtgtacaagt atgaaatgtc
      841 gtttcgaaca tatagatgtc tcgcaaacgt aaatatttat cgattgtcat aaaactttga
      901 ccttgtgaag tgtcaacctt gactgtcgaa ccaccatagt ttggcgcgaa ttgagcgtca
      961 taattgttta ctctcagtgc agtcaacatg tcgagtttcg tgccgaataa agagcaaacg
     1021 cggacagtat taattttctg ttttcatttg aagaaaacag ctgcggccgg atccaggaaa
     1081 cagctatgac catgattacg ccaagctacg taatacgact cactagtggg cagatcttcg
     1141 aatgcatcaa aagtgtctta tctagatttt taaaaatttg ttcttctcca cttgttctct
     1201 taataaaata aattctaaat tttgaacact aattagtgca ctaatcaatg ttttgaattt
     1261 tgaataaaat aattttaaaa ttttaaatgt tgtaaatttt tagtgatact gtaggtacaa
     1321 ggaaactatt ttattgaatt cttgatcaaa gaaggtaggc agtttatata aataatctta
     1381 ttgttagata ccattttgaa tatatgtttc agttccagaa aaataagcta tacatagtat
     1441 cgatttggaa catatataac gaaaacagac gtcatgaaat cacacattgg aaaagtgaga
     1501 aagttgttga gtagaggagg tgacaaaagt gagatgccaa ctcacttatc cattctactc
     1561 gttttgtgct cttttccact ttctgatgtc tgctcatctt cttattcttc aagcgttttg
     1621 tctattctca cacctctcca atctttcagt ttctgttttt aatgtctaga cacgcgtgta
     1681 acgtacaaca taaatgtttg acgtcaacca gatgtagtac aataaaaaat atattcagat
     1741 tacctagaaa attacttcgg tcatgaccac caattttttg atttttcgat tttcgaaacc
     1801 gacatcgttg aagaacattc aggtcataat tttttatttg aaaaataatg ttttctatgt
     1861 attaattcgg tttagtcgat cgaatatgta cagttgagaa ttactagagt acggtgcgca
     1921 agtatgcaaa cttgcgtaac ggtgtttaca catttgtcac gtttctctgc gtctcttcgt
     1981 ctcaacaatg cttgcactcc gtcacatttt cgaattgtcg gccgatatcg gtatttagat
     2041 aaattcagtg ctctcctttt tttctccatt ttactttcat caatcttttt taaaaatcaa
     2101 tattcagcaa actattgaaa aattaattca aaaaacacga ctggtcggaa gttatttgca
     2161 taatcaatag aaatttcgtt ttccgctcgc tgtcagtgtt ttgtcctgtt tgtatttccc
     2221 agtgcacttt tcgagtggct caagttttta tctattttcc tgcaaacttg catatattgt
     2281 tttcttccag aagcctttga tatctcaata attttatact tgtttcagaa atcgagcaaa
     2341 aaccaaggta tgtttcgaat gatactaata ataacttcgt atagcataca ttatacgaag
     2401 ttataacata acatagaaca ttttcagatg gtctcaaagg gtgaagaaga taacatggca
     2461 attattaaag agtttatgcg tttcaaggtg catatggagg gatctgtcaa tgggcatgag
     2521 tttgaaattg aaggtgaagg agaaggccga ccatatgagg gaacacaaac cgcaaaacta
     2581 aaggtaagtt taaacatata tatactaact aaccctgatt atttaaattt tcaggtaact
     2641 aaaggcggac cattaccatt cgcctgggac atcctctctc cacagttcat gtatggaagt
     2701 aaagcttatg ttaaacatcc ggcagatata ccagattatt tgaaactttc attcccggag
     2761 ggttttaagt gggaacgcgt aatgaatttt gaagacggag gagttgttac agtgacgcaa
     2821 gactcaaggt aagtttaaac agttcggtac taactaacca tacatattta aattttcagc
     2881 ctccaagatg gagaatttat ttataaagtc aaacttcgag gaacgaattt cccctcggat
     2941 ggacctgtta tgcagaagaa gactatggga tgggaagctt caagtgaaag aatgtaccct
     3001 gaagacggtg ctcttaaggg agagattaaa caacgtctta aattgaaaga tggaggacat
     3061 tacgatgctg aggtaagttt aaacatgatt ttactaacta actaatctga tttaaatttt
     3121 caggtgaaga caacttacaa agccaaaaaa ccagttcagc tgccaggagc gtacaatgtt
     3181 aatattaaac tggatatcac ctcccacaac gaggattaca ctatcgttga gcaatatgaa
     3241 agagctgaag ggcggcactc gacaggtggc atggatgaat tgtataagta gggatgatcg
     3301 acgccgacgt cgttgaattt tcaaatttta aatactgaat atttgttttt tttcctatta
     3361 tttatttatt ctctttgtgt tttttttctt gctttctaaa aaattaattc aatccaaatc
     3421 taaacatttt tttttctctt tccgtctccc aattcgtatt ccgctcctct catctgaaca
     3481 caatgtgcaa gtttatttat cttctcgctt tcatttcatt aggacgtggg gggaattggt
     3541 ggaaggggga aacacacaaa aggatgatgg aaatgaaata aggacacaca atatgcaaca
     3601 acattcaatt cagaaatatg gaggaaggtt taaaagaaaa cataaaaata tatagaggag
     3661 gaaggaaaac taaggtaagt ttaaactttc tcatactaat aataacttcg tatagcatac
     3721 attatacgaa gttatattaa ctaacgcgct ctatttaaat tttcagatga gtaaaggaga
     3781 agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca
     3841 caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa
     3901 atttatttgc actactggaa aactacctgt tccatgggta agtttaaaca tatatatact
     3961 aactaaccct gattatttaa attttcagcc aacacttgtc actactttct gttatggtgt
     4021 tcaatgcttc tcgagatacc cagatcatat gaaacggcat gactttttca agagtgccat
     4081 gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac
     4141 acgtaagttt aaacagttcg gtactaacta accatacata tttaaatttt caggtgctga
     4201 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     4261 agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata
     4321 catcatggca gacaaacaaa agaatggaat caaagttgta agtttaaaca tgattttact
     4381 aactaactaa tctgatttaa attttcagaa cttcaaaatt agacacaaca ttgaagatgg
     4441 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     4501 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     4561 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     4621 tgaactatac aaatagatgc aagatccttt caagcattcc cttcttctct atcactcttc
     4681 tttctttttg tcaaaaaatt ctctcgctaa tttatttgct tttttaatgt tattatttta
     4741 tgacttttta tagtcactga aaagtttgca tctgagtgaa gtgaatgcta tcaaaatgtg
     4801 attctgtctg atgtactttc acaatctctc ttcaattcca ttttgaagtg ctttaaaccc
     4861 gaaaggttga gaaaaatgcg agcgctcaaa tatttgtatt gtgttcgttg agtgacccaa
     4921 caaaaagagg aaactttatt gtgccgccaa gaaaaaagtc tcctgcagga tatctggatc
     4981 cacgaagctt cccatggtga cgtcaccggt tctagatacc taggtgagct ctggtaccct
     5041 ctagtcaagg ccttaagtga gtcgtattac ggactggccg tcgttttaca ggccggatgg
     5101 gccatggcac cttaacattt ctaaatttta gtttaataaa aaaattttaa aataatacga
     5161 gttttgagac taactttcaa taaaaaccaa aaaattgctt caaaaaagag ctcgcgccga
     5221 tttacgataa acacagcgcg ctacgacact ccgaaaaccg cgccgcacca ttacattctc
     5281 cgccccaccg cacggtttgc ggaaggcgat gaaattgctg ggcctaaaat tttaaccctt
     5341 tttttcccgt ttttcaactg ttttcccgat tttttcgata tttttttcag gtttttagtt
     5401 agaaaagtat ataaaaattg cattatttgt agttttcgct cgtaatttgt gatagaaaac
     5461 ggccaatttg cctagatttg tgattctacc aagtggaatt tttgggaaaa tatcaatatt
     5521 ccacgatttt tattcaaatc ataacgtttc gaaattatca gcttaaataa aactaaaata
     5581 aataataaaa tactaaaaac cctgaatttc tcggttctcg gttcagcttg agttcgttct
     5641 atacacgtta tcgatttttg tcatttcgtt taatcatggg ttttgattca ttttaaatat
     5701 gaaatttgat ttattcctcg tttcgaagtt ggtttgtctt gtccggccgg taaacaaatg
     5761 ttattaaaaa aaataataaa tttcgaaatt tttcgtaatg ttccgcaatc gtgtttttcc
     5821 gtttatctcg tcttcccttc cttcaaatat actatttatt ttctcaattc acatattttc
     5881 agccagccct gctcaacaag tggaataaaa atgattgaac aagatggatt gcacgcaggt
     5941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6061 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     6121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcacc agccgaactg
     6421 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtcca attactcttc
     6721 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
     6781 aacttctctt aatttctttg ttttttagct tcttttaagc cacctctaac aatgaaattg
     6841 tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc
     6901 ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt
     6961 tttttacttc tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt
     7021 atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc
     7081 tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa
     7141 tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg
     7201 tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg
     7261 atgcaagaaa gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt
     7321 gataatttga aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc
     7381 ccaatatacc aaacataact gtttctcgag gtcgacggta tcgataagct tgatccggat
     7441 ctgcggctta ctcaccagac ctggccccat ccgattacca cctattcgct tcgatgggac
     7501 acgcactcgc tgagcagcgc ttcgattctt acgaaagtgt gaaaaaatgg ctcgatgaat
     7561 ggttcgccgc aaaagacgat gagttctact ggcgtggaat ccacaaattg cccgagagat
     7621 gggaaaaatg tgtagctagc gacggcaaat actttgaata aatgattttt tctttttcca
     7681 caaaatttaa cgtgtttttt gatttaaaaa aaacgacatt tcatacttgt acacctgata
     7741 tctagatacc taggtgagct ctggtacccc cctggcgtta cccaacttaa tcgccttgca
     7801 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     7861 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     7921 ctgtgcggta tttcacaccg catatggtgc actctcagtg caatctgctc tgatgccgca
     7981 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     8041 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     8101 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     8161 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     8221 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     8281 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     8341 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     8401 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     8461 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     8521 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     8581 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     8641 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     8701 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     8761 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     8821 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     8881 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     8941 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     9001 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     9061 gcagcactgg ggtcagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     9121 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     9181 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     9241 ttttaattta aaaggatcta ggtgaagatc ctttttgata atc
//
