LOCUS       sequence-212577-        8205 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 8205)
  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 8205)
  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..8205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     286..303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    420..441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(456..461,462..479,480..486)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    494..510
                     /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     499..521
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     518..534
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     518..534
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4680..5390
                     /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(4825..4843)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5093..5112
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6308..6330)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6430..6449
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6527..6545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6613..6717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6718..6786,6787..7578)
                     /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(6936..6955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7749..8205,1..132)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
       61 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      301 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      361 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      421 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
      481 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
      541 tacgccaagc tgtaagttta aacatgatct tactaactaa ctattctcat ttaaattttc
      601 agagcttaaa aatggctgaa atcactcaca acgatggata cgctaacaac ttggaaatga
      661 aataagcttg ggctgcagcc cgggaaacat gatatcgacg gacatatttt tgattgatta
      721 tcataatttt cgtgggaatc ttcggaacat gctctttgcg gtttataagt ggagatgcgc
      781 caacgggaaa cttgttttaa aactattcct atggtgctaa catcaccaca tgaaaacttt
      841 tcaaaacttt tttgggtttt ttattttttt attattattc gtatctaaac attgtaggcg
      901 ttgtgatgtt gccctaaatt tctttaaaaa gagcgaacat tttcaaaaat ttgctattta
      961 aatcgaatga aaatttccat tcaaatattt gaaaaaaatt atgaaaaaat tccttttgtt
     1021 gcatcttcat cttctatgaa agttacaatt ttagttggat cagaaatatg agatatcaaa
     1081 aaaaaattaa gatttttttg tttgtttttt tgaagcctaa aatattttga tatcatttat
     1141 ctcttatgtc ttttgtttta tcgaaacgtt tacaacacat caactatgcg tcaataaatt
     1201 ttttaggtag ttaaacaaat gttgaaaaaa aaagcaaaag caaccaagag ctgttaaaat
     1261 agagaaaatg cagttatttt ttatattcaa gaaaagttat cacagaagtt ccgtaatgag
     1321 ttttttcctt atcaacatgt tcccatcaac ttcttagttt ctcctcattc ccattgttca
     1381 acataattat cacctctgac cttattctgc tctttttaag cacaggtgtt aatatgccgt
     1441 ccatcatatt tgccctccgt ctgccgcttt ccttatctga cctttacatc gcctttttgt
     1501 atatgtacat aataattgta tcatcatcaa cctccgccat aaacatgcat gtctttttac
     1561 ctcttttggc atgtaccgtt cttctttgta tttttattat gtcttccttc catcttccct
     1621 cttttttacg gattttcgga ttgcggaaga agaagatggt cgataattac cgtaatgagg
     1681 taatttgaaa catattggac agaagaaata cgcgcggatg ctttcggtgg caaggttaat
     1741 tcgttgtctt tgtttctgga atgtccttca taaaaaacac ggattaccac ctgtttttaa
     1801 cgtggattaa gatttttgga aacaatttac tctgcctaat ttttttgtac atttaaaaat
     1861 gtagacttac cgaattcacc gagcttgacc agttttaaag gttaccgttc acactaaaaa
     1921 ttgttgaaaa aattttcgac taaacctaat gattttattt taaaaaataa atgaaatagc
     1981 atacaaattt ggattttttt caattttttt caagaaaatc atgaaacatt tagaaaactc
     2041 gagtaacaag ctttccgttc attctgtaat ttttaagttc caaatttaaa aaaatgcatt
     2101 ttgatttact gggattgccg tgctcgtaat tatcagcaaa acatttgccg aattagccga
     2161 gctcgacaat ctattttttc gcaaactttt gccatgtttc gtaaaacaat tttccatttt
     2221 atggtagaca tttttcatca atgaattcac aaaatttaaa atccccactg aaagtaccaa
     2281 aatcttcctg aaaaatccgg caaaaaatcc atctcattat catgtcacat atacatgttc
     2341 tattggcact atgtttggga tgtcctccaa ttaatacttc tgtgtcaatc tcatcggcgc
     2401 ccggcggcca ctaccaaatg caaaactttg tttgtaaaag aagaagccaa taattttctc
     2461 gttattcatt tttattactc tttttgcgtt tcattcttct ttgttcgaat tgatgaacgc
     2521 acgttgcttc tctctggttt tcctgccgtt ccttatattg ccaaggaaac caatgcggat
     2581 tttcggaaga ttttcttcat gtcctttaga aaatatcacc acttttcttc tttttccatt
     2641 tcaaatttca atttccatac aactcatctt ctattatagc ttttcaggtg ttcgactgtt
     2701 tgaagataag aatcaattaa aatttggttc actgagcatg tcatccactt ttccttttct
     2761 ttgttctaaa attcaccaaa ctgaattttc cttaaccgga aatttacatg aaaataagaa
     2821 acatggatta catcctttac atatcatttt tgttttttta tgtgtattcc atgttcccaa
     2881 aagtttgatg caatttttga ttctttttca attttccaaa ttcgtgtctc gcagggaacg
     2941 attcacagtc caacattccc aaattgcgaa attttagaat atatattccc aagcgtcgaa
     3001 tatctcaaaa atttgtcgtt ttctcagttt gacatgtcac ctttgccggt tcctctattc
     3061 gtctattagc tctccagaga gcaatgtcat gtcacacgaa gaattcaggg acatgtttcg
     3121 tttttttttg tttgcatgtg gtcaccgata tgggaacttg aactttccaa tttttgcttg
     3181 ctttttcttt tccggaaact taatactatt tgagaaactc cgcatcttat ttgtttgatc
     3241 ccaaattctg aaattcctct ttgaatttga tctcaagctt cactataaaa aattgtttca
     3301 ggaaaattgg aagagcaagc ggatccccgg gattggccaa aggacccaaa ggtatgtttc
     3361 gaatgatact aacataacat agaacatttt caggaggacc cttggctagc aaaaatgagt
     3421 ttcccggtgg catttttgct cgttctcttc ttggttccat tcacatttgc gacaaataat
     3481 ttagttgtta gacagaaaga attggaaatt gtaattggag gagaagtctc tgttaatttc
     3541 cagatcaaaa accacacatc acaatcattg aacgcaacac gtatctccct ctcgcagtca
     3601 ccatacatct cccatcctga tgctattctc gttgacaatt ggaatgctaa tgtgacagtt
     3661 ttaggaagcc aattggtgtc cggagctatt ttagaagctc tcaattgtac tacagatggg
     3721 tcaattactt gtcctctgga tcttgaagat gcgtttgcta gaatcacagt tattcgctct
     3781 cactttcttg ccattttgat tcaaattgtt ggatggacat acttcttcgc atggagtatt
     3841 tcgttttatc cccaaatgta tctgaatttc aaaagaaaaa gcgtcgtcgg gctaaacttc
     3901 gatttcttat ctctgaacct tgtcggattt tgtgcatatg ctattttcaa tctgttaatg
     3961 tattataata gtcatgtaaa gaatgaatac aatattgtga atcctcggtc accgccacca
     4021 gtacttttga acgatgttgt gttcgctgtt cacgcatttc ttgcctgctt tatcacaatt
     4081 cttcaatgtt tattctatga acgtgataat cagagtgttt cttcaaaatg tatcgccctt
     4141 atgattgtgc tgatctcttt tggattctgc tccgctgccg ccacggttct caggaaaatt
     4201 caattgctct cgtttgtgac cagtttgtcg tacataaaaa tggctgtaac ctgctgtaaa
     4261 tatttcccac aggcttactt caactatacc cgaaaaagta cagttggatg gtcaatcgga
     4321 aacatcatgc tagatttcac aggaggaact ctcgatattc ttcaaatgat tcttcaagca
     4381 gttaatgtga acgattggtc tgcattttat gcgaatcccg tcaaattcgg actgggattt
     4441 gtctcaatat tctttgacat cattttcatg gttcaacatt atgtattgta tccaaatgca
     4501 gaagttccac acaatgaata tcatggtgtt gacaatccca atccagacaa tattgctcgt
     4561 gatgctgaac aatatgccgg cgacagtgaa tcgatggagt caacggaacc tattattgta
     4621 catgacaccg gtcaggtgag tataattgta caatgtaagt tttaaatgta aatttttaga
     4681 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     4741 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     4801 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     4861 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     4921 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     4981 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     5041 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     5101 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     5161 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     5221 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     5281 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     5341 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa atgaattcca
     5401 actgagcgcc ggtcgctacc attaccaact tgtctggtgt caaaaataat aggggccgct
     5461 gtcatcagag taagtttaaa ctgagttcta ctaactaacg agtaatattt aaattttcag
     5521 catctcgcgc ccgtgcctct gacttctaag tccaattact cttcaacatc cctacatgct
     5581 ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaacttc tcttaatttc
     5641 tttgtttttt agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata
     5701 gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat
     5761 tctatcccaa aatctacaca atgttctgtg tacacttctt atgtttttta cttctgataa
     5821 atttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata tgttacgttt
     5881 cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga cgtcaaatca
     5941 tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt gaaagtttat
     6001 gaaattaatt ttcctgcttt tgctttttgg ggtttcccct attgtttgtc aagatttcga
     6061 ggacggcgtt tttcttgcta aaatcacaag tattgatgag cacgatgcaa gaaagatcgg
     6121 aagaaggttt gggtttgagg ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg
     6181 agtagtgtct atggggtttt tgccttaaat gacagaatac attcccaata taccaaacat
     6241 aactgtttcc tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg
     6301 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     6361 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     6421 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     6481 cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc
     6541 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6601 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6661 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6721 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6781 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6841 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6901 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6961 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7021 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7081 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     7141 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7201 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7261 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7321 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7381 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7441 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7501 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7561 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7621 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7681 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7741 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7801 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7861 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7921 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7981 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     8041 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     8101 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     8161 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtc
//
