LOCUS       sequence-208430-        8210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the lysosome marker CTNS-1-mCherry cDNA in a subset of 
            ventral cord cholinergic motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8210)
  AUTHORS   Edwards SL, Yu SC, Hoover CM, Phillips BC, Richmond JE, Miller KG
  TITLE     An organelle gatekeeper function for Caenorhabditis elegans UNC-16 
            (JIP3) at the axon initial segment.
  JOURNAL   Genetics. 2013 May;194(1):143-61. doi: 10.1534/genetics.112.147348.
  PUBMED    23633144
REFERENCE   2  (bases 1 to 8210)
  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"; doi: 
            "10.1534/genetics.112"; journalName: "Genetics"; date: "2013-05"; 
            volume: "194"; issue: "1"; pages: "143-61"
COMMENT     Sequence Label: KG#371
FEATURES             Location/Qualifiers
     source          1..8210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4017..4727
                     /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(4162..4180)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4430..4449
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5650..5672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5772..5791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5869..5887)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5955..6059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6060..6128,6129..6920)
                     /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(6278..6297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7091..7679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7580..7599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7833..7850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7967..7988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8003..8008,8009..8026,8027..8033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8041..8057
                     /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     8046..8068
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8065..8081
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8065..8081
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcagcccgg gaaacatgat atcgacggac atatttttga ttgattatca
       61 taattttcgt gggaatcttc ggaacatgct ctttgcggtt tataagtgga gatgcgccaa
      121 cgggaaactt gttttaaaac tattcctatg gtgctaacat caccacatga aaacttttca
      181 aaactttttt gggtttttta tttttttatt attattcgta tctaaacatt gtaggcgttg
      241 tgatgttgcc ctaaatttct ttaaaaagag cgaacatttt caaaaatttg ctatttaaat
      301 cgaatgaaaa tttccattca aatatttgaa aaaaattatg aaaaaattcc ttttgttgca
      361 tcttcatctt ctatgaaagt tacaatttta gttggatcag aaatatgaga tatcaaaaaa
      421 aaattaagat ttttttgttt gtttttttga agcctaaaat attttgatat catttatctc
      481 ttatgtcttt tgttttatcg aaacgtttac aacacatcaa ctatgcgtca ataaattttt
      541 taggtagtta aacaaatgtt gaaaaaaaaa gcaaaagcaa ccaagagctg ttaaaataga
      601 gaaaatgcag ttatttttta tattcaagaa aagttatcac agaagttccg taatgagttt
      661 tttccttatc aacatgttcc catcaacttc ttagtttctc ctcattccca ttgttcaaca
      721 taattatcac ctctgacctt attctgctct ttttaagcac aggtgttaat atgccgtcca
      781 tcatatttgc cctccgtctg ccgctttcct tatctgacct ttacatcgcc tttttgtata
      841 tgtacataat aattgtatca tcatcaacct ccgccataaa catgcatgtc tttttacctc
      901 ttttggcatg taccgttctt ctttgtattt ttattatgtc ttccttccat cttccctctt
      961 ttttacggat tttcggattg cggaagaaga agatggtcga taattaccgt aatgaggtaa
     1021 tttgaaacat attggacaga agaaatacgc gcggatgctt tcggtggcaa ggttaattcg
     1081 ttgtctttgt ttctggaatg tccttcataa aaaacacgga ttaccacctg tttttaacgt
     1141 ggattaagat ttttggaaac aatttactct gcctaatttt tttgtacatt taaaaatgta
     1201 gacttaccga attcaccgag cttgaccagt tttaaaggtt accgttcaca ctaaaaattg
     1261 ttgaaaaaat tttcgactaa acctaatgat tttattttaa aaaataaatg aaatagcata
     1321 caaatttgga tttttttcaa tttttttcaa gaaaatcatg aaacatttag aaaactcgag
     1381 taacaagctt tccgttcatt ctgtaatttt taagttccaa atttaaaaaa atgcattttg
     1441 atttactggg attgccgtgc tcgtaattat cagcaaaaca tttgccgaat tagccgagct
     1501 cgacaatcta ttttttcgca aacttttgcc atgtttcgta aaacaatttt ccattttatg
     1561 gtagacattt ttcatcaatg aattcacaaa atttaaaatc cccactgaaa gtaccaaaat
     1621 cttcctgaaa aatccggcaa aaaatccatc tcattatcat gtcacatata catgttctat
     1681 tggcactatg tttgggatgt cctccaatta atacttctgt gtcaatctca tcggcgcccg
     1741 gcggccacta ccaaatgcaa aactttgttt gtaaaagaag aagccaataa ttttctcgtt
     1801 attcattttt attactcttt ttgcgtttca ttcttctttg ttcgaattga tgaacgcacg
     1861 ttgcttctct ctggttttcc tgccgttcct tatattgcca aggaaaccaa tgcggatttt
     1921 cggaagattt tcttcatgtc ctttagaaaa tatcaccact tttcttcttt ttccatttca
     1981 aatttcaatt tccatacaac tcatcttcta ttatagcttt tcaggtgttc gactgtttga
     2041 agataagaat caattaaaat ttggttcact gagcatgtca tccacttttc cttttctttg
     2101 ttctaaaatt caccaaactg aattttcctt aaccggaaat ttacatgaaa ataagaaaca
     2161 tggattacat cctttacata tcatttttgt ttttttatgt gtattccatg ttcccaaaag
     2221 tttgatgcaa tttttgattc tttttcaatt ttccaaattc gtgtctcgca gggaacgatt
     2281 cacagtccaa cattcccaaa ttgcgaaatt ttagaatata tattcccaag cgtcgaatat
     2341 ctcaaaaatt tgtcgttttc tcagtttgac atgtcacctt tgccggttcc tctattcgtc
     2401 tattagctct ccagagagca atgtcatgtc acacgaagaa ttcagggaca tgtttcgttt
     2461 ttttttgttt gcatgtggtc accgatatgg gaacttgaac tttccaattt ttgcttgctt
     2521 tttcttttcc ggaaacttaa tactatttga gaaactccgc atcttatttg tttgatccca
     2581 aattctgaaa ttcctctttg aatttgatct caagcttcac tataaaaaat tgtttcagga
     2641 aaattggaag agcaagcgga tccccgggat tggccaaagg acccaaaggt atgtttcgaa
     2701 tgatactaac ataacataga acattttcag gaggaccctt ggctagcaaa aatgagtttc
     2761 ccggtggcat ttttgctcgt tctcttcttg gttccattca catttgcgac aaataattta
     2821 gttgttagac agaaagaatt ggaaattgta attggaggag aagtctctgt taatttccag
     2881 atcaaaaacc acacatcaca atcattgaac gcaacacgta tctccctctc gcagtcacca
     2941 tacatctccc atcctgatgc tattctcgtt gacaattgga atgctaatgt gacagtttta
     3001 ggaagccaat tggtgtccgg agctatttta gaagctctca attgtactac agatgggtca
     3061 attacttgtc ctctggatct tgaagatgcg tttgctagaa tcacagttat tcgctctcac
     3121 tttcttgcca ttttgattca aattgttgga tggacatact tcttcgcatg gagtatttcg
     3181 ttttatcccc aaatgtatct gaatttcaaa agaaaaagcg tcgtcgggct aaacttcgat
     3241 ttcttatctc tgaaccttgt cggattttgt gcatatgcta ttttcaatct gttaatgtat
     3301 tataatagtc atgtaaagaa tgaatacaat attgtgaatc ctcggtcacc gccaccagta
     3361 cttttgaacg atgttgtgtt cgctgttcac gcatttcttg cctgctttat cacaattctt
     3421 caatgtttat tctatgaacg tgataatcag agtgtttctt caaaatgtat cgcccttatg
     3481 attgtgctga tctcttttgg attctgctcc gctgccgcca cggttctcag gaaaattcaa
     3541 ttgctctcgt ttgtgaccag tttgtcgtac ataaaaatgg ctgtaacctg ctgtaaatat
     3601 ttcccacagg cttacttcaa ctatacccga aaaagtacag ttggatggtc aatcggaaac
     3661 atcatgctag atttcacagg aggaactctc gatattcttc aaatgattct tcaagcagtt
     3721 aatgtgaacg attggtctgc attttatgcg aatcccgtca aattcggact gggatttgtc
     3781 tcaatattct ttgacatcat tttcatggtt caacattatg tattgtatcc aaatgcagaa
     3841 gttccacaca atgaatatca tggtgttgac aatcccaatc cagacaatat tgctcgtgat
     3901 gctgaacaat atgccggcga cagtgaatcg atggagtcaa cggaacctat tattgtacat
     3961 gacaccggtc aggtgagtat aattgtacaa tgtaagtttt aaatgtaaat ttttagatgg
     4021 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4081 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4141 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4201 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4261 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4321 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     4381 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     4441 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     4501 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     4561 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     4621 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     4681 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagaa ttccaactga
     4741 gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat
     4801 cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct
     4861 cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc
     4921 tccctgtgct cccaccccct atttttgtta ttatcaaaaa aacttcttct taatttcttt
     4981 gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa
     5041 ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct
     5101 atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttttac ttctgataaa
     5161 ttttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt
     5221 cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca
     5281 tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat
     5341 gaaattaatt ttcctgcttt tgctttttgg gggtttcccc tattgtttgt caagagtttc
     5401 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc
     5461 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt
     5521 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac
     5581 ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt tcggtgatga
     5641 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5701 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5761 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5821 accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac
     5881 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5941 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6001 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6061 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6121 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6181 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6241 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6301 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6361 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     6421 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     6481 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     6541 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     6601 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6661 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6721 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6781 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6841 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6901 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6961 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7021 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7081 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7141 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7201 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7261 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7321 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7381 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7441 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc
     7501 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7561 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7621 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7681 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7741 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7801 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7861 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7921 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7981 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     8041 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctgt
     8101 aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga gcttaaaaat
     8161 ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
