LOCUS       sequence-373426-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone with ORF encoding NLS lacZ flanked by Gateway 
            recombination sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  AUTHORS   Roure A, Rothbacher U, Robin F, Kalmar E, Ferone G, Lamy C, Missero 
            C, Mueller F, Lemaire P
  TITLE     A multicassette Gateway vector set for high throughput and 
            comparative analyses in ciona and vertebrate embryos.
  JOURNAL   PLoS One. 2007. 2(9):e916.
  PUBMED    17878951
REFERENCE   2  (bases 1 to 5757)
  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: "PLoS One. 
            2007. 2(9):e916."
COMMENT     Sequence Label: pENT-L1-Kozak-NLSlacZ-stop-L2
FEATURES             Location/Qualifiers
     source          1..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(281..308)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(400..486)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     549..566
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     550..566
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    582..681
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             715..735
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             823..3870
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     protein_bind    complement(3895..3994)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4011..4030)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4012..4030)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4035..4051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4035..4051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4153..4172)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4164..4973
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4231..4250)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5120..5708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5609..5628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
       61 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      121 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      181 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      241 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
      301 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
      361 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
      421 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
      481 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
      541 cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt
      601 gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa
      661 ctttgtacaa aaaagcaggc tgcaagggta ccgagctcag aaaaaatgac tgctccaaag
      721 aagaagcgta aggtaccggt gggtgaagac cagaaacagc acctcgaact gagccgcgat
      781 attgcccagc gtttcaacgc gctgtatggc gagatcgatc ccgtcgtttt acaacgtcgt
      841 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
      901 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
      961 aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg
     1021 gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt
     1081 tacgatgcgc ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt
     1141 cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta
     1201 caggaaggcc agacgcgaat tatttttgat ggcgttaact cggcgtttca tctgtggtgc
     1261 aacgggcgct gggtcggtta cggccaggac agtcgtttgc cgtctgaatt tgacctgagc
     1321 gcatttttac gcgccggaga aaaccgcctc gcggtgatgg tgctgcgctg gagtgacggc
     1381 agttatctgg aagatcagga tatgtggcgg atgagcggca ttttccgtga cgtctcgttg
     1441 ctgcataaac cgactacaca aatcagcgat ttccatgttg ccactcgctt taatgatgat
     1501 ttcagccgcg ctgtactgga ggctgaagtt cagatgtgcg gcgagttgcg tgactaccta
     1561 cgggtaacag tttctttatg gcagggtgaa acgcaggtcg ccagcggcac cgcgcctttc
     1621 ggcggtgaaa ttatcgatga gcgtggtggt tatgccgatc gcgtcacact acgtctgaac
     1681 gtcgaaaacc cgaaactgtg gagcgccgaa atcccgaatc tctatcgtgc ggtggttgaa
     1741 ctgcacaccg ccgacggcac gctgattgaa gcagaagcct gcgatgtcgg tttccgcgag
     1801 gtgcggattg aaaatggtct gctgctgctg aacggcaagc cgttgctgat tcgaggcgtt
     1861 aaccgtcacg agcatcatcc tctgcatggt caggtcatgg atgagcagac gatggtgcag
     1921 gatatcctgc tgatgaagca gaacaacttt aacgccgtgc gctgttcgca ttatccgaac
     1981 catccgctgt ggtacacgct gtgcgaccgc tacggcctgt atgtggtgga tgaagccaat
     2041 attgaaaccc acggcatggt gccaatgaat cgtctgaccg atgatccgcg ctggctaccg
     2101 gcgatgagcg aacgcgtaac gcgaatggtg cagcgcgatc gtaatcaccc gagtgtgatc
     2161 atctggtcgc tggggaatga atcaggccac ggcgctaatc acgacgcgct gtatcgctgg
     2221 atcaaatctg tcgatccttc ccgcccggtg cagtatgaag gcggcggagc cgacaccacg
     2281 gccaccgata ttatttgccc gatgtacgcg cgcgtggatg aagaccagcc cttcccggct
     2341 gtgccgaaat ggtccatcaa aaaatggctt tcgctacctg gagagacgcg cccgctgatc
     2401 ctttgcgaat acgcccacgc gatgggtaac agtcttggcg gtttcgctaa atactggcag
     2461 gcgtttcgtc agtatccccg tttacagggc ggcttcgtct gggactgggt ggatcagtcg
     2521 ctgattaaat atgatgaaaa cggcaacccg tggtcggctt acggcggtga ttttggcgat
     2581 acgccgaacg atcgccagtt ctgtatgaac ggtctggtct ttgccgaccg cacgccgcat
     2641 ccagcgctga cggaagcaaa acaccagcag cagtttttcc agttccgttt atccgggcaa
     2701 accatcgaag tgaccagcga atacctgttc cgtcatagcg ataacgagct cctgcactgg
     2761 atggtggcgc tggatggtaa gccgctggca agcggtgaag tgcctctgga tgtcgctcca
     2821 caaggtaaac agttgattga actgcctgaa ctaccgcagc cggagagcgc cgggcaactc
     2881 tggctcacag tacgcgtagt gcaaccgaac gcgaccgcat ggtcagaagc cgggcacatc
     2941 agcgcctggc agcagtggcg tctggcggaa aacctcagtg tgacgctccc cgccgcgtcc
     3001 cacgccatcc cgcatctgac caccagcgaa atggattttt gcatcgagct gggtaataag
     3061 cgttggcaat ttaaccgcca gtcaggcttt ctttcacaga tgtggattgg cgataaaaaa
     3121 caactgctga cgccgctgcg cgatcagttc acccgtgcac cgctggataa cgacattggc
     3181 gtaagtgaag cgacccgcat tgaccctaac gcctgggtcg aacgctggaa ggcggcgggc
     3241 cattaccagg ccgaagcagc gttgttgcag tgcacggcag atacacttgc tgatgcggtg
     3301 ctgattacga ccgctcacgc gtggcagcat caggggaaaa ccttatttat cagccggaaa
     3361 acctaccgga ttgatggtag tggtcaaatg gcgattaccg ttgatgttga agtggcgagc
     3421 gatacaccgc atccggcgcg gattggcctg aactgccagc tggcgcaggt agcagagcgg
     3481 gtaaactggc tcggattagg gccgcaagaa aactatcccg accgccttac tgccgcctgt
     3541 tttgaccgct gggatctgcc attgtcagac atgtataccc cgtacgtctt cccgagcgaa
     3601 aacggtctgc gctgcgggac gcgcgaattg aattatggcc cacaccagtg gcgcggcgac
     3661 ttccagttca acatcagccg ctacagtcaa cagcaactga tggaaaccag ccatcgccat
     3721 ctgctgcacg cggaagaagg cacatggctg aatatcgacg gtttccatat ggggattggt
     3781 ggcgacgact cctggagccc gtcagtatcg gcggaattcc agctgagcgc cggtcgctac
     3841 cattaccagt tggtctggtg tcaaaaataa taataaccgg gcaggccatg tctgacccag
     3901 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     3961 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     4021 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     4081 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     4141 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     4201 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     4261 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     4321 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     4381 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     4441 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     4501 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     4561 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     4621 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     4681 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     4741 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     4801 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     4861 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     4921 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     4981 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     5041 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     5101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggcctt
//
