LOCUS       sequence-223062-        6380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector to produce humanized Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6380)
  AUTHORS   Pillai RS, Bhattacharyya SN, Artus CG, Zoller T, Cougot N, Basyuk E,
            Bertrand E, Filipowicz W
  TITLE     Inhibition of translational initiation by Let-7 MicroRNA in human 
            cells.
  JOURNAL   Science. 2005 Sep 2. 309(5740):1573-6.
  PUBMED    16081698
REFERENCE   2  (bases 1 to 6380)
  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: "Science. 
            2005 Sep 2. 309(5740):1573-6."
COMMENT     Sequence Label: p CIneo-RL
FEATURES             Location/Qualifiers
     source          1..6380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        800..1179
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1180..1391
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1342..1362
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1388..1412
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1552..1684
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1667..1686
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1729..1747
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1751..1760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1757..2692
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2746..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2757..2776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2811..2830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3053..3508
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3140..3159)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3350..3371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3625..3982
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3625..3816
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3648..3668)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3833..3968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4033..4827
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4087..4106)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4697..4716
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4891..4939
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4980..4999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5099..5121
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5159..5177)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5245..5349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5350..5418,5419..6210)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5568..5587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
      661 cttcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
      721 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
      781 ccaatatgac cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg
      841 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      901 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      961 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1021 gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat
     1081 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact
     1141 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     1201 accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     1261 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     1321 tgcgatcgcc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1381 taagcagagc tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc
     1441 acagttaaat tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca
     1501 gtgactctct taaggtagcc ttgcagaagt tggtcgtgag gcactgggca ggtaagtatc
     1561 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     1621 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     1681 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
     1741 ctataggcta gccaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat
     1801 cactgggcct cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa
     1861 ctactatgat tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc
     1921 ctccagctac ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat
     1981 ccctgatctg atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct
     2041 ggatcactac aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat
     2101 ctttgtgggc cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga
     2161 caagatcaag gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga
     2221 gtggcctgac atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt
     2281 gcttgagaat aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga
     2341 gcctgaggag ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc
     2401 taccctctcc tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca
     2461 gattgtccgc aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat
     2521 cgagtccgac cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa
     2581 caccgagttc gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat
     2641 gggtaagtac atcaagagct tcgtggagcg cgtgctgaag aacgagcagt aattctagag
     2701 cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg
     2761 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2821 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2881 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     2941 acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga
     3001 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     3061 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3121 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3181 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3241 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3301 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3361 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3421 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3481 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     3541 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     3601 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     3661 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3721 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3841 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3901 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3961 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     4021 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4081 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4141 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4201 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4261 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4321 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4381 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4441 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4501 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4561 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4621 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4681 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4741 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4801 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4861 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     4921 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt
     4981 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5041 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5101 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5161 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5221 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5281 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5341 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5401 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5461 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5521 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5581 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5641 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5701 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5761 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5821 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5881 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5941 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6001 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6061 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6121 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6181 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6241 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6301 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6361 gaaaagatca aaggatcttc
//
