LOCUS       sequence-403000-        6370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate2-tag fused CDC-42 with 8xMS2(V1) stem loops in C. 
            elegans, and the tissue-specific promoter col-19 is used..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6370)
  AUTHORS   Hu Y, Xu J, Gao E, Fan X, Wei J, Ye B, Xu S, Ma W
  TITLE     Enhanced single RNA imaging reveals dynamic gene expression in live 
            animals.
  JOURNAL   Elife. 2023 Mar 3;12:e82178. doi: 10.7554/eLife.82178.
  PUBMED    36867026
REFERENCE   2  (bases 1 to 6370)
  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: "Elife."; 
            date: "2023-03-3"; volume: "12:e82178. doi"; pages: " 
            10.7554/eLife.82178"
COMMENT     Sequence Label: pSX2739
FEATURES             Location/Qualifiers
     source          1..6370
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     299..320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        483..587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(588..656,657..1448)
                     /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(806..825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1619..2207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2108..2127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2361..2378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2495..2516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2531..2536,2537..2554,2555..2561)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2569..2585
                     /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     2574..2596
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2593..2609
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2593..2609
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2628..2648
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2630..2648
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    4359..4383
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5020..5038
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5059..5077
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5104..5122
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5143..5161
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5188..5206
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5227..5245
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5272..5290
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        5311..5329
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     protein_bind    complement(5364..5388)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6185..6204)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6186..6204)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6214..6231)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6214..6230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6223..6245)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      421 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
      481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      541 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      601 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      661 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      721 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      781 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      841 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      901 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      961 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1021 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1081 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1141 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1201 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1261 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1321 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1381 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1441 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1741 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2281 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2341 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2401 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2461 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2521 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2581 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     2641 actaaaggga acaaaagctg gagctccatt tgaaaatttg caccaatgta ttatttaatt
     2701 ttttttttcg aaaatttaac gcattttctc tctaaaaact cgaaatttag tgtgttctaa
     2761 acaacagtaa gcatacaata cctttgttca aaattgacgt gctttctgaa ccaatatggt
     2821 tagtttcaaa aatttttgta ttataggata gaaatatttg gaaataaatt ttaaaaccaa
     2881 acttatgcct ttctctttta gtatcccagc taggtaatat tttagtattt gcccaaatcc
     2941 ttgaagtaag gagtatataa tttttgaaaa acaataaaac tccagataat tcatagtttt
     3001 ttctcgaaag aaaatttttg agattagtta ttgaacttca tttttgaaca ttattcgttg
     3061 aaaaacactc gctttgtctt tattttcaaa aaaattccga tttccccaac cagaaaaaaa
     3121 aacagataga agaaatttct ccttaaattt cattgtccat ctctcttgga aacacattat
     3181 ctatcaaatg aaaaacgcat tttttttcct ggcagaaaaa tgaaattggt tagattacac
     3241 tggttaggtt tgaaggtgta actttcgctt tctcagcaac tttcagtata aaaggaaacg
     3301 gtcaccattt agaaagacat cagttcatca actcgaggca tgcctgcagg tcgactctag
     3361 aggatccccg ggattggcca aaggacccaa aggtatgttt cgaatgatac taacataaca
     3421 tagaacattt tcaggaggac ccttggaggg tacaggtaga gctcaccggt atgtccgagc
     3481 tcatcaagga gaacatgcac atgaagctct acatggaggg aaccgtcaac aaccaccact
     3541 tcaagtgcac ctccgaggga gagggaaagc catacgaggg aacccaaacc atgcgtatca
     3601 aggtaagttt aaacatatat atactaacta accctgatta tttaaatttt caggccgtcg
     3661 agggaggacc actcccattc gccttcgaca tcctcgccac ctccttcatg tacggatcca
     3721 agaccttcat caaccacacc caaggaatcc cagacttctt caagcaatcc ttcccagagg
     3781 gattcacctg ggagcgtgtc accacctacg aggacggagg agtcctcacc gccacccaag
     3841 acacctccct ccaagacgga tgcctcatct acaacgtcaa ggtaagttta aacagttcgg
     3901 tactaactaa ccatacatat ttaaattttc agatccgtgg agtcaacttc ccatccaacg
     3961 gaccagtcat gcaaaagaag accctcggat gggaggcctc caccgagacc ctctacccag
     4021 ccgacggagg actcgaggga cgtgccgaca tggccctcaa gctcgtcgga ggaggacacc
     4081 tcatctgcaa cctcaaggta agtttaaaca tgattttact aactaactaa tctgatttaa
     4141 attttcagac cacctaccgt tccaagaagc cagccaagaa cctcaagatg ccaggagtct
     4201 actacgtcga ccgtcgtctc gagcgtatca aggtggccga caaggagacc tacgtcgagc
     4261 aacacgaggt cgccgtcgcc cgttactgcg acctcccatc caagctcgga caccgtaccg
     4321 gtactagtgg atcccccggg ctgcaggaat tcgatatcac aagtttgtac aaaaaagcag
     4381 gctccgaatt cgcccttatg cagacgatca agtgcgtcgt cgttggagat ggagctgtcg
     4441 gtaaaacttg tctcctgatc agctatacca caaacaagtt tccttctgag tatgtgccga
     4501 cagtcttcga caattacgcc gtcacagtaa tgatcggtgg cgagccatac acattaggat
     4561 tgtttgatac tgctggacag gaagattacg atcgattaag gcctctatcg tatccacaga
     4621 ccgacgtgtt tcttgtttgc ttctccgtgg ttgctccagc ttcattcgag aatgtccgag
     4681 aaaaatgggt gcctgaaatt tcgcatcatt gctcaaagac cccattcttg ttagttggta
     4741 ctcaagtcga tctcagggat gatccaggaa tgctcgagaa actggcaaaa aacaagcaga
     4801 aaccagtgtc aacggatgtt ggagagaagt tggcaaagga attgaaagca gtgaaatacg
     4861 ttgaatgctc agcgttgacg cagaagggac tgaaaaatgt attcgacgaa gccattctgg
     4921 ccgctctcga cccaccacaa caggagaaga agaagaagtg caatattctc taggggaggt
     4981 gatagcattg cttggatccc tgcaggtcga ctctagaaaa catgaggatc acccatgtct
     5041 gcaggtcgac tctagaaaac atgaggatca cccatgtgaa ttcctgcagg tcgactctag
     5101 aaaacatgag gatcacccat gtctgcaggt cgactctaga aaacatgagg atcacccatg
     5161 tgatatcctg caggtcgact ctagaaaaca tgaggatcac ccatgtctgc aggtcgactc
     5221 tagaaaacat gaggatcacc catgtctcga gctgcaggtc gactctagaa aacatgagga
     5281 tcacccatgt ctgcaggtcg actctagaaa acatgaggat cacccatgtg ggcccgttta
     5341 aacccgctga aagggcgaat tcgacccagc tttcttgtac aaagtggtga tatcaagctt
     5401 atcgattgtc atcagatcgc catctcgcgc ccgtgcctct gacttctaag tccaattact
     5461 cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc
     5521 aaaaaacttc tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa
     5581 attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc
     5641 ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt
     5701 atgtttttta cttctgataa atttttttga aacatcatag aaaaaaccgc acacaaaata
     5761 ccttatcata tgttacgttt cagtttatga ccgcaatttt tatttcttcg cacgtctggg
     5821 cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt ttggaatttt
     5881 tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt tgctttttgg ggtttcccct
     5941 attgtttgtc aagatttcga ggacggcgtt tttcttgcta aaatcacaag tattgatgag
     6001 cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga
     6061 agttgataat ttgaaagtgg agtagtgtct atggggtttt tgccttaaat gacagaatac
     6121 attcccaata taccaaacat aactgtttct actagtcggc cgtacgggcc cggtacccaa
     6181 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     6241 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     6301 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     6361 tggcgaatgg
//
