LOCUS       sequence-12131-e        5997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector containing LacZ..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5997)
  AUTHORS   Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja 
            R, Thomas SR, Alkan O, Bhimdi T, Green TM, Johannessen CM, Silver 
            SJ, Nguyen C, Murray RR, Hieronymus H, Balcha D, Fan C, Lin C, 
            Ghamsari L, Vidal M, Hahn WC, Hill DE, Root DE
  TITLE     A public genome-scale lentiviral expression library of human ORFs.
  JOURNAL   Nat Methods. 2011 Jun 26;8(8):659-61. doi: 10.1038/nmeth.1638
  PUBMED    21706014
REFERENCE   2  (bases 1 to 5997)
  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: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-06-26- 26"; volume: "8"; 
            issue: "8"; pages: "659-61"
COMMENT     Sequence Label: pDONR223_LacZ
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             683..3727
                     /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
                     SWSPSVSAELQLSAGRYHYQLVWCQK"
     protein_bind    complement(3747..3771)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3824..3865
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(3876..3975)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3992..4011)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3993..4011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4016..4032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4016..4032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4463..5254
                     /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      5347..5935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5836..5855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgatagatc ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      721 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      781 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc
      841 ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc
      901 cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac
      961 caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg
     1021 ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat
     1081 tatttttgat ggcgttaact cggcgtttca tctgtggtgc aacgggcgct gggtcggtta
     1141 cggccaggac agtcgtttgc cgtctgaatt tgacctgagc gcatttttac gcgccggaga
     1201 aaaccgcctc gcggtgatgg tgctgcgctg gagtgacggc agttatctgg aagatcagga
     1261 tatgtggcgg atgagcggca ttttccgtga cgtctcgttg ctgcataaac cgactacaca
     1321 aatcagcgat ttccatgttg ccactcgctt taatgatgat ttcagccgcg ctgtactgga
     1381 ggctgaagtt cagatgtgcg gcgagttgcg tgactaccta cgggtaacag tttctttatg
     1441 gcagggtgaa acgcaggtcg ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga
     1501 gcgtggtggt tatgccgatc gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg
     1561 gagcgccgaa atcccgaatc tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac
     1621 gctgattgaa gcagaagcct gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct
     1681 gctgctgctg aacggcaagc cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc
     1741 tctgcatggt caggtcatgg atgagcagac gatggtgcag gatatcctgc tgatgaagca
     1801 gaacaacttt aacgccgtgc gctgttcgca ttatccgaac catccgctgt ggtacacgct
     1861 gtgcgaccgc tacggcctgt atgtggtgga tgaagccaat attgaaaccc acggcatggt
     1921 gccaatgaat cgtctgaccg atgatccgcg ctggctaccg gcgatgagcg aacgcgtaac
     1981 gcgaatggtg cagcgcgatc gtaatcaccc gagtgtgatc atctggtcgc tggggaatga
     2041 atcaggccac ggcgctaatc acgacgcgct gtatcgctgg atcaaatctg tcgatccttc
     2101 ccgcccggtg cagtatgaag gcggcggagc cgacaccacg gccaccgata ttatttgccc
     2161 gatgtacgcg cgcgtggatg aagaccagcc cttcccggct gtgccgaaat ggtccatcaa
     2221 aaaatggctt tcgctacctg gagagacgcg cccgctgatc ctttgcgaat acgcccacgc
     2281 gatgggtaac agtcttggcg gtttcgctaa atactggcag gcgtttcgtc agtatccccg
     2341 tttacagggc ggcttcgtct gggactgggt ggatcagtcg ctgattaaat atgatgaaaa
     2401 cggcaacccg tggtcggctt acggcggtga ttttggcgat acgccgaacg atcgccagtt
     2461 ctgtatgaac ggtctggtct ttgccgaccg cacgccgcat ccagcgctga cggaagcaaa
     2521 acaccagcag cagtttttcc agttccgttt atccgggcaa accatcgaag tgaccagcga
     2581 atacctgttc cgtcatagcg ataacgagct cctgcactgg atggtggcgc tggatggtaa
     2641 gccgctggca agcggtgaag tgcctctgga tgtcgctcca caaggtaaac agttgattga
     2701 actgcctgaa ctaccgcagc cggagagcgc cgggcaactc tggctcacag tacgcgtagt
     2761 gcaaccgaac gcgaccgcat ggtcagaagc cggccacatc agcgcctggc agcagtggcg
     2821 tctggcggaa aacctcagtg tgacgctccc cgccgcgtcc cacgccatcc cgcatctgac
     2881 caccagcgaa atggattttt gcatcgagct gggtaataag cgttggcaat ttaaccgcca
     2941 gtcaggcttt ctttcacaga tgtggattgg cgataaaaaa caactgctga cgccgctgcg
     3001 cgatcagttc acccgtgcac cgctggataa cgacattggc gtaagtgaag cgacccgcat
     3061 tgaccctaac gcctgggtcg aacgctggaa ggcggcgggc cattaccagg ccgaagcagc
     3121 gttgttgcag tgcacggcag atacacttgc tgacgcggtg ctgattacga ccgctcacgc
     3181 gtggcagcat caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag
     3241 tggtcaaatg gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg
     3301 gattggcctg aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg
     3361 gccgcaagaa aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc
     3421 attgtcagac atgtataccc cgtacgtctt cccgagcgaa aacggtctgc gctgcgggac
     3481 gcgcgaattg aattatggcc cacaccagtg gcgcggcgac ttccagttca acatcagccg
     3541 ctacagtcaa cagcaactga tggaaaccag ccatcgccat ctgctgcacg cggaagaagg
     3601 cacatggctg aatatcgacg gtttccatat ggggattggt ggagacgact cctggagccc
     3661 gtcagtatcg gcggaattac agctgagcgc cggtcgctac cattaccagt tggtctggtg
     3721 tcaaaaaact aagggtgggc gcgccgaccc agctttcttg tacaaagtgg ttgatatcca
     3781 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3841 ccctctcctc ggtctcgatt ctacgcgtac cggtttgcca actttcttgt acaaagttgg
     3901 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     3961 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     4021 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     4081 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
     4141 gctgcccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     4201 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     4261 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     4321 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     4381 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     4441 ccctaaaaca aagttaaaca tcatgaggga agcggtgatc gccgaagtat cgactcaact
     4501 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     4561 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     4621 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     4681 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     4741 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     4801 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     4861 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     4921 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     4981 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     5041 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     5101 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     5161 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     5221 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     5281 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5461 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5701 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
