LOCUS       sequence-178093-        3980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) 
            Renilla Luciferase CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3980)
  AUTHORS   Jackel C, Nogueira MS, Ehni N, Kraus C, Ranke J, Dohmann M, Noessner
            E, Nelson PJ
  TITLE     A vector platform for the rapid and efficient engineering of stable 
            complex transgenes.
  JOURNAL   Sci Rep. 2016 Oct 3;6:34365. doi: 10.1038/srep34365.
  PUBMED    27694838
REFERENCE   2  (bases 1 to 3980)
  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: "Sci Rep."; 
            date: "2016-10-3"; volume: "6:34365. doi"; pages: " 
            10.1038/srep34365"
COMMENT     Sequence Label: pENTR221-Renilla
FEATURES             Location/Qualifiers
     source          1..3980
                     /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    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          683..815
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     798..817
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        860..878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             888..1823
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1854..1975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1865..1884)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1919..1938
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(2092..2191)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2222..2240)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2245..2261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2245..2261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2363..2382)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2374..3183
                     /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(2441..2460)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3330..3918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3819..3838
                     /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 tgtatacaaa
      661 agttgtatga ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca
      721 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
      781 tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag
      841 ctcttaaggc tagagtactt aatacgactc actataggct agccaccatg acttcgaaag
      901 tttatgatcc agaacaaagg aaacggatga taactggtcc gcagtggtgg gccagatgta
      961 aacaaatgaa tgttcttgat tcatttatta attattatga ttcagaaaaa catgcagaaa
     1021 atgctgttat ttttttacat ggtaacgcgg cctcttctta tttatggcga catgttgtgc
     1081 cacatattga gccagtagcg cggtgtatta taccagacct tattggtatg ggcaaatcag
     1141 gcaaatctgg taatggttct tataggttac ttgatcatta caaatatctt actgcatggt
     1201 ttgaacttct taatttacca aagaagatca tttttgtcgg ccatgattgg ggtgcttgtt
     1261 tggcatttca ttatagctat gagcatcaag ataagatcaa agcaatagtt cacgctgaaa
     1321 gtgtagtaga tgtgattgaa tcatgggatg aatggcctga tattgaagaa gatattgcgt
     1381 tgatcaaatc tgaagaagga gaaaaaatgg ttttggagaa taacttcttc gtggaaacca
     1441 tgttgccatc aaaaatcatg agaaagttag aaccagaaga atttgcagca tatcttgaac
     1501 cattcaaaga gaaaggtgaa gttcgtcgtc caacattatc atggcctcgt gaaatcccgt
     1561 tagtaaaagg tggtaaacct gacgttgtac aaattgttag gaattataat gcttatctac
     1621 gtgcaagtga tgatttacca aaaatgttta ttgaatcgga cccaggattc ttttccaatg
     1681 ctattgttga aggtgccaag aagtttccta atactgaatt tgtcaaagta aaaggtcttc
     1741 atttttcgca agaagatgca cctgatgaaa tgggaaaata tatcaaatcg ttcgttgagc
     1801 gagttctcaa aaatgaacaa taattctaga gcggccgctt cgagcagaca tgataagata
     1861 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     1921 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     1981 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2041 caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccaggtggca aacccagctt
     2101 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     2161 tcactatcag tcaaaataaa atcattattt gccatccagc tgcagggcgg ccgcgatatc
     2221 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2281 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     2341 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     2401 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     2461 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     2521 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     2581 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     2641 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     2701 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     2761 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     2821 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     2881 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     2941 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     3001 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     3061 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     3121 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     3181 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     3241 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     3301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3961 tggccttttg ctcacatgtt
//
