LOCUS       sequence-257699-        5989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Marker of microtubule dynamics in C. elegans hypodermis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5989)
  AUTHORS   Chuang M, Hsiao TI, Tong A, Xu S, Chisholm AD
  TITLE     DAPK interacts with Patronin and the microtubule cytoskeleton in 
            epidermal development and wound repair.
  JOURNAL   Elife. 2016 Sep 23;5. doi: 10.7554/eLife.15833.
  PUBMED    27661253
REFERENCE   2  (bases 1 to 5989)
  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: "2016-09-23"; volume: "5. doi"; pages: " 10.7554/eLife.15833"
COMMENT     Sequence Label: pCZGY1584 Pcol-19::ebp-2::mGFP::unc-54 3'UTR
FEATURES             Location/Qualifiers
     source          1..5989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1114..1138)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1161..1163,1164..1166,1167..1880)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1206..1227)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1467..1486)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1814..1835
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2706..2728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2828..2847
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2925..2943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3011..3115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3116..3184,3185..3976)
                     /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(3334..3353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4147..4735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4636..4655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4889..4906
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5023..5044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5059..5064,5065..5082,5083..5089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5097..5113
                     /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     5102..5124
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5121..5137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5121..5137
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    join(5982..5989,1..17)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gtacaaaaaa gcaggctccg aattcgccct tgttttcaga aaccgaggca aaatggtcgt
       61 caacgtgttc atctcggctg tgaccactga cacgctgagc cggaaggaag cagttgcttg
      121 ggtcaataac ctactcaagg tttgaatttg cgggaaatgt tcaaattatt taatatttca
      181 aatttcagag tcacttcacc aaagtggaag agatggcgtc tggagccgcg tactgtcagc
      241 tcactcatct cttgttcaac gccatcaacc tgaagaaggt caaattcaac ccacggagcg
      301 agcccgatgt tctgaacaat tggaaggtgc tcacaacgac ctggaaggat ctcggcattg
      361 acaagccggt cgatgtcgag aaaatgaaga aggccaagtt ccaggacaac atggagtttt
      421 tgcaatggtt ctacaaggta cgttcttttt gtgagaggca taataacttt ttaataactt
      481 tttttttgca gttctacaat gcgaacctga ccacggagcc agaagaatat gatgcagttg
      541 gtgctcgatt cggagaggac ttgcccgctc tgaaaggatc cacaggctct cgtccagttg
      601 ctgctcctgc tagaccggtg gctgccgctc cgccaaagcc aaggcctgct gcgccagcgg
      661 ttgcggctcc agctgtgaaa gcatcagttc caccgacagt tcgcaacggc acccggcccg
      721 ctccagcgac atcagccgga actcgacgcg ctgacccatc tgcaagcgaa gaacttctca
      781 aacaaggttt gagattccgt gataattgta ctattattaa atttttgttt ttagaaatcg
      841 agaaacacaa ggccgccagt gatgaatggg agacgacggc caaggaaatg gagacagaac
      901 gagaatacta ctactcaatt cttcagcgtg tcgaaagtct tgccaacgaa gcagaggaga
      961 gtggttccag tacggtcgac gtggcggctc tgaagacgat tctctacgcc ggaaacgagg
     1021 acactgagca ggtcgatgaa atcgaaaacg aacatcttgt cgagtctctg caggcaaatc
     1081 tggacgatac ggagactttc aagggcgaat tcgacccagc tttcttgtac aaagtggtga
     1141 tgtaccatgg tattgatatc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1201 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1261 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1321 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1381 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1441 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1501 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1561 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1621 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1681 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1741 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     1801 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1861 tggacgagct gtacaagtaa gatatctgag ctccgcatcg gccgctgtca tcagatcgcc
     1921 atctcgcgcc cgtgcctctg acttctaagt ccaattactc ttcaacatcc ctacatgctc
     1981 tttctccctg tgctcccacc ccctattttt gttattatca aaaaacttct cttaatttct
     2041 ttgtttttta gcttctttta agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag
     2101 aattaattcg taataaaaag tcgaaaaaaa ttgtgctccc tccccccatt aataataatt
     2161 ctatcccaaa atctacacaa tgttctgtgt acacttctta tgttttttac ttctgataaa
     2221 tttttttgaa acatcataga aaaaaccgca cacaaaatac cttatcatat gttacgtttc
     2281 agtttatgac cgcaattttt atttcttcgc acgtctgggc ctctcatgac gtcaaatcat
     2341 gctcatcgtg aaaaagtttt ggagtatttt tggaattttt caatcaagtg aaagtttatg
     2401 aaattaattt tcctgctttt gctttttggg gtttccccta ttgtttgtca agatttcgag
     2461 gacggcgttt ttcttgctaa aatcacaagt attgatgagc acgatgcaag aaagatcgga
     2521 agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa gttgataatt tgaaagtgga
     2581 gtagtgtcta tggggttttt gccttaaatg acagaataca ttcccaatat accaaacata
     2641 actgtttcta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2701 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2761 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2821 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2881 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     2941 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3001 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3061 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3121 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3181 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3241 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3301 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3361 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3421 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3481 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3541 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3601 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3661 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3721 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3781 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3841 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3901 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3961 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4261 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4861 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4921 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4981 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5041 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5101 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttttttt
     5161 ctacctgtac cctccaaggg tcctcctgaa aatgttctat gttatgttag tatcattcga
     5221 aacatacctt tgggtccttt ggccaatccc ggggatcctc tagagtcgac ctgcaggcat
     5281 gcctcgagtt gatgaactga tgtctttcta aatggtgacc gtttcctttt atactgaaag
     5341 ttgctgagaa agcgaaagtt acaccttcaa acctaaccag tgtaatctaa ccaatttcat
     5401 ttttctgcca ggaaaaaaaa tgcgtttttc atttgataga taatgtgttt ccaagagaga
     5461 tggacaatga aatttaagga gaaatttctt ctatctgttt ttttttctgg ttggggaaat
     5521 cggaattttt ttgaaaataa agacaaagcg agtgtttttc aacgaataat gttcaaaaat
     5581 gaagttcaat aactaatctc aaaaattttc tttcgagaaa aaactatgaa ttatctggag
     5641 ttttattgtt tttcaaaaat tatatactcc ttacttcaag gatttgggca aatactaaaa
     5701 tattacctag ctgggatact aaaagagaaa ggcataagtt tggttttaaa atttatttcc
     5761 aaatatttct atcctataat acaaaaattt ttgaaactaa ccatattggt tcagaaagca
     5821 cgtcaatttt gaacaaaggt attgtatgct tactgttgtt tagaacacac taaatttcga
     5881 gtttttagag agaaaatgcg ttaaattttc gaaaaaaaaa attaaataat acattggtgc
     5941 aaattttcaa atgctggtac cgctagcgtc gacggtacat cacaagttt
//
