LOCUS       sequence-214098-        6128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pmec-4 Gateway Destination vector for cloning for expression in C. 
            elegans mechanosensory neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6128)
  AUTHORS   Ghosh-Roy A, Wu Z, Goncharov A, Jin Y, Chisholm AD
  TITLE     Calcium and cyclic AMP promote axonal regeneration in Caenorhabditis
            elegans and require DLK-1 kinase.
  JOURNAL   J Neurosci. 2010 Mar 3;30(9):3175-83. doi: 
            10.1523/JNEUROSCI.5464-09.2010.
  PUBMED    20203177
REFERENCE   2  (bases 1 to 6128)
  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: 
            "10.1523/JNEUROSCI.5464-09"; journalName: "J Neurosci"; date: 
            "2010-03-3- 3"; volume: "30"; issue: "9"; pages: "3175-83"
COMMENT     Sequence Label: pCZGY553
FEATURES             Location/Qualifiers
     source          1..6128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1156..1280
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1305..1310,1311..1328,1329..1335)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1389..2048
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1455..1474)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2390..2695
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2574..2593
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2736..2860)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3702..3724)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3824..3843
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3921..3939)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4007..4111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4112..4180,4181..4972)
                     /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(4330..4349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5143..5731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5632..5651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5885..5902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6019..6040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6055..6060,6061..6078,6079..6085)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6093..6109
                     /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     6098..6120
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6117..6128,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6117..6128,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttcaataca agctcaaata caatactatt ttttttccgt
       61 ctgaacataa aattagtttc taaataaaaa atgtgaaaat aaatatttgg ttttaatcgt
      121 taaaatcttc caaattacta ataaagttca gtcaactata tgcttcgaaa ttcactgaat
      181 ctattattgt tattaatttt tagacccttg gagagctccc aactctttca atccagaaga
      241 ctgatggatg ccaggtgtac ttgtcaaagg tggctcaagg atgcgagatt gtcacttcca
      301 aatccagtga gatgaacatc tccgtgcaaa ccaatgatga tggggattat gtaagttttt
      361 cggattgtag aacccaatct aaatttcctt ttcagtcgga gttcccggtt ccggagcagt
      421 tcaagacaac ttttgtcaat ggcaagctag tcactgtcgt ctcggacatt tgctaaatct
      481 tctcacgtca taaccttatt tttcaacttc atgaactatc ccctcgaact tttttatttg
      541 tccctgtcta caattcttta taagcaattg agacacactt tgctttatta aataataccg
      601 attttgtttt gtatatgcct tgactggtgc tcaactattt ttattgaaaa aattactttt
      661 tactgtctcg tcgaataaaa gtgatccctt ccaaaaaata gttattcatg tttgaaggtt
      721 tgtgatcgta gaacaattgc accgaatgta gtgtaaaaaa aattccagaa tgttcaaata
      781 catttccacc ctcaccagaa catctcaaac accaggcaca tttcaacaca ctttcatgga
      841 tcttatcttg cctcaaaacc atcagctttc ttctcattct acaagtatga aacaatctac
      901 tattctatac atttataatg taatggaacc atgtgaagac atgagcaaga tcattgtcgt
      961 cagcgactac ccctccttca tttttcttag agtatcagtt tgctaatgct atttttttta
     1021 tcgctatcaa gttatagagg atccccggga ttggccaaag gacccaaagg tatgtttcga
     1081 atgatactaa cataacatag aacattttca ggaggaccct tgcttggagg gtaccgctag
     1141 cgtcgacggt acatcacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata
     1201 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
     1261 caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt
     1321 ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg
     1381 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1441 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1501 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1561 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
     1621 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     1681 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     1741 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     1801 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     1861 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     1921 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1981 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2041 gggcgtaaac gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     2101 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     2161 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     2221 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     2281 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     2341 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     2401 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     2461 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     2521 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     2581 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     2641 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     2701 aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt
     2761 tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat
     2821 cattttacgt ttctcgttca gctttcttgt acaaagtggt gatgtaccat ggtattgata
     2881 tctgagctcc gcatcggccg ctgtcatcag atcgccatct cgcgcccgtg cctctgactt
     2941 ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct
     3001 atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagtc
     3061 acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga
     3121 aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt
     3181 ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa
     3241 accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt
     3301 cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag
     3361 tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt
     3421 tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc
     3481 acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag
     3541 tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct
     3601 taaatgacag aatacattcc caatatacca aacataactg tttctactag tcggccgtac
     3661 gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3721 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3781 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     3841 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3901 cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     3961 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4021 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4081 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4141 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4201 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4261 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4321 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4381 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4441 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4501 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4561 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4621 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4681 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4741 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4801 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4861 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4921 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4981 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5041 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5101 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5161 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5221 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5281 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5341 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5401 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5461 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5521 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5581 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5641 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5701 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5761 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5821 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5881 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5941 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6001 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6061 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6121 aaacagct
//
