LOCUS       sequence-170564-        3430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector for inserting promotors for tissue specific 
            expression of NLS-tdMCP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3430)
  AUTHORS   Campbell PD, Chao JA, Singer RH, Marlow FL
  TITLE     Dynamic visualization of transcription and RNA subcellular 
            localization in zebrafish.
  JOURNAL   Development. 2015 Apr 1;142(7):1368-74. doi: 10.1242/dev.118968. 
            Epub 2015 Mar 10.
  PUBMED    25758462
REFERENCE   2  (bases 1 to 3430)
  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: 
            "10.1242/dev.118968"; journalName: "Development"; date: "2015-04-1- 
            1"; volume: "142"; issue: "7"; pages: "1368-74"
COMMENT     Sequence Label: pME-NLS-tdMCP
FEATURES             Location/Qualifiers
     source          1..3430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(36..63)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(155..241)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     290..312
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     304..321
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     305..321
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    337..436
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             617..637
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             641..667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             641..667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             668..679
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1077..1176)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1193..1212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1194..1212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1217..1233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1217..1233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1664..2455
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2548..3136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3037..3056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3290..3307
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
       61 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
      121 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
      181 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
      241 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
      301 cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg
      361 atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg
      421 tacaaaaaag caggctccga attcgccctt gtcccttctc ggcgattctg cggagggatc
      481 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
      541 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
      601 ggcggccgca atgggcccaa aaaagaaaag aaaagttggc tacccctacg acgtgcccga
      661 ctacgccatc gaaggccgcc atatgctagc cgttaaaatg gcttctaact ttactcagtt
      721 cgttctcgtc gacaatggcg gaactggcga cgtgactgtc gccccaagca acttcgctaa
      781 cgggatcgct gaatggatca gctctaactc gcgttcacag gcttacaaag taacctgtag
      841 cgttcgtcag agctctgcgc agaatcgcaa atacaccatc aaagtcgagg tgcctaaagg
      901 cgcctggcgt tcgtacttaa atatggaact aaccattcca attttcgcca cgaattccga
      961 ctgcgagctt attgttaagg caatgcaagg tctcctaaaa gatggaaacc cgattccctc
     1021 agcaatcgca gcaaactccg gcatctacgc ggattctaga atgaagggcg aattcgaccc
     1081 agctttcttg tacaaagttg gcattataaa aaataattgc tcatcaattt gttgcaacga
     1141 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     1201 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     1261 ctgatgttac attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga
     1321 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     1381 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     1441 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     1501 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     1561 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     1621 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
     1681 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     1741 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     1801 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     1861 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     1921 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     1981 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     2041 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     2101 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     2161 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     2221 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     2281 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     2341 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     2401 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     2461 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3421 aattaatacg
//
