LOCUS       sequence-373427-        5690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone with ORF encoding lacZ flanked by Gateway recombination 
            sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5690)
  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 5690)
  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-lacZ-stop-L2
FEATURES             Location/Qualifiers
     source          1..5690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..106
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(265..292)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(384..470)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     533..550
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     534..550
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    566..665
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             764..3811
                     /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(3812..3911)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3928..3947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3929..3947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3952..3968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3952..3968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4070..4089)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4081..4890
                     /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(4148..4167)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5037..5625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5526..5545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
       61 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      121 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      181 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
      241 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
      301 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
      361 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      421 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
      481 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
      541 gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg
      601 ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc
      661 aggctcagaa aaaatgccgg tgggtgaaga ccagaaacag cacctcgaac tgagccgcga
      721 tattgcccag cgtttcaacg cgctgtatgg cgagatcgat cccgtcgttt tacaacgtcg
      781 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      841 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
      901 gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct
      961 ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg
     1021 ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt
     1081 tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct
     1141 acaggaaggc cagacgcgaa ttatttttga tggcgttaac tcggcgtttc atctgtggtg
     1201 caacgggcgc tgggtcggtt acggccagga cagtcgtttg ccgtctgaat ttgacctgag
     1261 cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct ggagtgacgg
     1321 cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg acgtctcgtt
     1381 gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct ttaatgatga
     1441 tttcagccgc gctgtactgg aggctgaagt tcagatgtgc ggcgagttgc gtgactacct
     1501 acgggtaaca gtttctttat ggcagggtga aacgcaggtc gccagcggca ccgcgccttt
     1561 cggcggtgaa attatcgatg agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa
     1621 cgtcgaaaac ccgaaactgt ggagcgccga aatcccgaat ctctatcgtg cggtggttga
     1681 actgcacacc gccgacggca cgctgattga agcagaagcc tgcgatgtcg gtttccgcga
     1741 ggtgcggatt gaaaatggtc tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt
     1801 taaccgtcac gagcatcatc ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca
     1861 ggatatcctg ctgatgaagc agaacaactt taacgccgtg cgctgttcgc attatccgaa
     1921 ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa
     1981 tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc gctggctacc
     2041 ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat
     2101 catctggtcg ctggggaatg aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg
     2161 gatcaaatct gtcgatcctt cccgcccggt gcagtatgaa ggcggcggag ccgacaccac
     2221 ggccaccgat attatttgcc cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc
     2281 tgtgccgaaa tggtccatca aaaaatggct ttcgctacct ggagagacgc gcccgctgat
     2341 cctttgcgaa tacgcccacg cgatgggtaa cagtcttggc ggtttcgcta aatactggca
     2401 ggcgtttcgt cagtatcccc gtttacaggg cggcttcgtc tgggactggg tggatcagtc
     2461 gctgattaaa tatgatgaaa acggcaaccc gtggtcggct tacggcggtg attttggcga
     2521 tacgccgaac gatcgccagt tctgtatgaa cggtctggtc tttgccgacc gcacgccgca
     2581 tccagcgctg acggaagcaa aacaccagca gcagtttttc cagttccgtt tatccgggca
     2641 aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc gataacgagc tcctgcactg
     2701 gatggtggcg ctggatggta agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc
     2761 acaaggtaaa cagttgattg aactgcctga actaccgcag ccggagagcg ccgggcaact
     2821 ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat
     2881 cagcgcctgg cagcagtggc gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc
     2941 ccacgccatc ccgcatctga ccaccagcga aatggatttt tgcatcgagc tgggtaataa
     3001 gcgttggcaa tttaaccgcc agtcaggctt tctttcacag atgtggattg gcgataaaaa
     3061 acaactgctg acgccgctgc gcgatcagtt cacccgtgca ccgctggata acgacattgg
     3121 cgtaagtgaa gcgacccgca ttgaccctaa cgcctgggtc gaacgctgga aggcggcggg
     3181 ccattaccag gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt
     3241 gctgattacg accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa
     3301 aacctaccgg attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag
     3361 cgatacaccg catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg
     3421 ggtaaactgg ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg
     3481 ttttgaccgc tgggatctgc cattgtcaga catgtatacc ccgtacgtct tcccgagcga
     3541 aaacggtctg cgctgcggga cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga
     3601 cttccagttc aacatcagcc gctacagtca acagcaactg atggaaacca gccatcgcca
     3661 tctgctgcac gcggaagaag gcacatggct gaatatcgac ggtttccata tggggattgg
     3721 tggcgacgac tcctggagcc cgtcagtatc ggcggaattc cagctgagcg ccggtcgcta
     3781 ccattaccag ttggtctggt gtcaaaaata aacccagctt tcttgtacaa agttggcatt
     3841 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     3901 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     3961 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     4021 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     4081 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     4141 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     4201 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4261 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4321 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4381 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     4441 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4501 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     4561 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4621 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4681 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     4741 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4801 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4861 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     4921 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     4981 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5041 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5101 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5161 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5221 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5281 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5341 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5401 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5461 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5521 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5581 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5641 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
//
