LOCUS       sequence-12130-e        3477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector containing HcRed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3477)
  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 3477)
  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_HcRed
FEATURES             Location/Qualifiers
     source          1..3477
                     /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     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ 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             671..1354
                     /codon_start=1
                     /product="far-red fluorescent variant of Heteractis crispa 
                     chromoprotein"
                     /label=HcRed1
                     /note="mammalian codon-optimized"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     protein_bind    complement(1356..1455)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1472..1491)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1473..1491)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1496..1512)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1496..1512)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1943..2734
                     /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      2827..3415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3316..3335
                     /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 atggtgagcg gcctgctgaa ggagagtatg cgcatcaaga tgtacatgga
      721 gggcaccgtg aacggccact acttcaagtg cgagggcgag ggcgacggca accccttcgc
      781 cggcacccag agcatgagaa tccacgtgac cgagggcgcc cccctgccct tcgccttcga
      841 catcctggcc ccctgctgcg agtacggcag caggaccttc gtgcaccaca ccgccgagat
      901 ccccgacttc ttcaagcaga gcttccccga gggcttcacc tgggagagaa ccaccaccta
      961 cgaggacggc ggcatcctga ccgcccacca ggacaccagc ctggagggca actgcctgat
     1021 ctacaaggtg aaggtgcacg gcaccaactt ccccgccgac ggccccgtga tgaagaacaa
     1081 gagcggcggc tgggagccca gcaccgaggt ggtgtacccc gagaacggcg tgctgtgcgg
     1141 ccggaacgtg atggccctga aggtgggcga ccggcacctg atctgccacc actacaccag
     1201 ctaccggagc aagaaggccg tgcgcgccct gaccatgccc ggcttccact tcaccgacat
     1261 ccggctccag atgctgcgga agaagaagga cgagtacttc gagctgtacg aggccagcgt
     1321 ggcccggtac agcgacctgc ccgagaaggc caacttgcca actttcttgt acaaagttgg
     1381 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1441 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     1501 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1561 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
     1621 gctgcccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     1681 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     1741 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     1801 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     1861 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     1921 ccctaaaaca aagttaaaca tcatgaggga agcggtgatc gccgaagtat cgactcaact
     1981 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     2041 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     2101 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     2161 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     2221 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     2281 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     2341 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     2401 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     2461 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     2521 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     2581 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     2641 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     2701 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     2761 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2941 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3181 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     3241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
