LOCUS       sequence-170470-        4380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector for inserting promotors for tissue specific 
            expression of NLS-tdMCP-tagRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4380)
  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 4380)
  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-NLStdMCP- tagRFP
FEATURES             Location/Qualifiers
     source          1..4380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1346..1445)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1462..1481)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1463..1481)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1486..1502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1486..1502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1933..2724
                     /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      2817..3405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3306..3325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3559..3576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3735..3762)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3854..3940)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3989..4011
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4003..4020
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4004..4020
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4036..4135
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4168..4188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4192..4218
                     /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             4192..4218
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4219..4230
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gtaacctgta gcgttcgtca gagctctgcg cagaatcgca aatacaccat caaagtcgag
       61 gtgcctaaag gcgcctggcg ttcgtactta aatatggaac taaccattcc aattttcgcc
      121 acgaattccg actgcgagct tattgttaag gcaatgcaag gtctcctaaa agatggaaac
      181 ccgattccct cagcaatcgc agcaaactcc ggcatctacg ccatggcttc taactttact
      241 cagttcgttc tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc
      301 gctaacggga tcgctgaatg gatcagctct aactcgcgtt cacaggctta caaagtaacc
      361 tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct
      421 aaaggcgcct ggcgttcgta cttaaatatg gaactaacca ttccaatttt cgccacgaat
      481 tccgactgcg agcttattgt taaggcaatg caaggtctcc taaaagatgg aaacccgatt
      541 ccctcagcaa tcgcagcaaa ctccggcatc tacgcggatt ctagaatggt gagcaagggc
      601 gaggagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtgaacaac
      661 caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
      721 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc
      781 ttcatgtacg gcagcagaac cttcatcaac cacacccagg gcatccccga cttctttaag
      841 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
      901 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc
      961 agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact cggctgggag
     1021 gccaacaccg agatgctgta ccccgctgac ggcggcctgg aaggcagaac cgacatggcc
     1081 ctgaagctcg tgggcggggg ccacctgatc tgcaacttca agaccacata cagatccaag
     1141 aaacccgcta agaacctcaa gatgcccggc gtctactatg tggaccacag actggaaaga
     1201 atcaaggagg ccgacaaaga gacctacgtc gagcagcacg aggtggctgt ggccagatac
     1261 tgcgacctcc ctagcaaact ggggcacaaa cttaatggca tggacgagct gtacaagtaa
     1321 accggttctc tcaagggcga attcgaccca gctttcttgt acaaagttgg cattataaaa
     1381 aataattgct catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1441 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1501 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1561 tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag
     1621 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc
     1681 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
     1741 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
     1801 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1861 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     1921 aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
     1981 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
     2041 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
     2101 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
     2161 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
     2221 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
     2281 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
     2341 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
     2401 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
     2461 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
     2521 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
     2581 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
     2641 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
     2701 atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg
     2761 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2821 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2881 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2941 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3001 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3061 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3121 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3181 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3241 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3301 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3361 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3421 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3481 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3541 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3601 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3661 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3721 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3781 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     3841 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     3901 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     3961 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     4021 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
     4081 caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccgaa
     4141 ttcgcccttg agagactcga gatgggccca aaaaagaaaa gaaaagttgg ctacccctac
     4201 gacgtgcccg actacgccat cgaaggccgc catatgctag ccgttaaaat ggcttctaac
     4261 tttactcagt tcgttctcgt cgacaatggc ggaactggcg acgtgactgt cgccccaagc
     4321 aacttcgcta acgggatcgc tgaatggatc agctctaact cgcgttcaca ggcttacaaa
//
