LOCUS       sequence-478462-        8023 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-LATS1 Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8023)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-LATS1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(5102..5120)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5157..5278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5168..5187)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5222..5241
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5523..5880
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5523..5714
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5546..5566)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5731..5866
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5895..5942
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5974..6768
                     /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(6028..6047)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6638..6657
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6832..6880
                     /note="synthetic polyadenylation signal"
     rep_origin      7037..7582
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     7483..7502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7699..7716
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat gaagaggagt gaaaagccag
     1621 aaggatatag acaaatgagg cctaagacct ttcctgccag taactatact gtcagtagcc
     1681 ggcaaatgtt acaagaaatt cgggaatccc ttaggaattt atctaaacca tctgatgctg
     1741 ctaaggctga gcataacatg agtaaaatgt caaccgaaga tcctcgacaa gtcagaaatc
     1801 cacccaaatt tgggacgcat cataaagcct tgcaggaaat tcgaaactct ctgcttccat
     1861 ttgcaaatga aacaaattct tctcggagta cttcagaagt taatccacaa atgcttcaag
     1921 acttgcaagc tgctggattt gatgaggata tggttataca agctcttcag aaaactaaca
     1981 acagaagtat agaagcagca attgaattca ttagtaaaat gagttaccaa gatcctcgac
     2041 gagagcagat ggctgcagca gctgccagac ctattaatgc cagcatgaaa ccagggaatg
     2101 tgcagcaatc agttaaccgc aaacagagct ggaaaggttc taaagaatcc ttagttcctc
     2161 agaggcatgg cccgccacta ggagaaagtg tggcctatca ttctgagagt cccaactcac
     2221 agacagatgt aggaagacct ttgtctggat ctggtatatc agcatttgtt caagctcacc
     2281 ctagcaacgg acagagagtg aaccccccac caccacctca agtaaggagt gttactcctc
     2341 caccacctcc aagaggccag actccccctc caagaggtac aactccacct cccccttcat
     2401 gggaaccaaa ctctcaaaca aagcgctatt ctggaaacat ggaatacgta atctcccgaa
     2461 tctctcctgt cccacctggg gcatggcaag agggctatcc tccaccacct ctcaacactt
     2521 cccccatgaa tcctcctaat caaggacaga gaggcattag ttctgttcct gttggcagac
     2581 aaccaatcat catgcagagt tctagcaaat ttaactttcc atcagggaga cctggaatgc
     2641 agaatggtac tggacaaact gatttcatga tacaccaaaa tgttgtccct gctggcactg
     2701 tgaatcggca gccaccacct ccatatcctc tgacagcagc taatggacaa agcccttctg
     2761 ctttacaaac agggggatct gctgctcctt cgtcatatac aaatggaagt attcctcagt
     2821 ctatgatggt gccaaacaga aatagtcata acatggaact atataacatt agtgtacctg
     2881 gactgcaaac aaattggcct cagtcatctt ctgctccagc ccagtcatcc ccgagcagtg
     2941 ggcatgaaat ccctacatgg caacctaaca taccagtgag gtcaaattct tttaataacc
     3001 cattaggaaa tagagcaagt cactctgcta attctcagcc ttctgctaca acagtcactg
     3061 caattacacc agctcctatt caacagcctg tgaaaagtat gcgtgtatta aaaccagagc
     3121 tacagactgc tttagcacct acacaccctt cttggatacc acagccaatt caaactgttc
     3181 aacccagtcc ttttcctgag ggaaccgctt caaatgtgac tgtgatgcca cctgttgctg
     3241 aagctccaaa ctatcaagga ccaccaccac cctacccaaa acatctgctg caccaaaacc
     3301 catctgttcc tccatacgag tcaatcagta agcctagcaa agaggatcag ccaagcttgc
     3361 ccaaggaaga tgagagtgaa aagagttatg aaaatgttga tagtggggat aaagaaaaga
     3421 aacagattac aacttcacct attactgtta ggaaaaacaa gaaagatgaa gagcgaaggg
     3481 aatctcgtat tcaaagttat tctcctcaag catttaaatt ctttatggag caacatgtag
     3541 aaaatgtact caaatctcat cagcagcgtc tacatcgtaa aaaacaatta gagaatgaaa
     3601 tgatgcgggt tggattatct caagatgccc aggatcaaat gagaaagatg ctttgccaaa
     3661 aagaatctaa ttacatccgt cttaaaaggg ctaaaatgga caagtctatg tttgtgaaga
     3721 taaagacact aggaatagga gcatttggtg aagtctgtct agcaagaaaa gtagatacta
     3781 aggctttgta tgcaacaaaa actcttcgaa agaaagatgt tcttcttcga aatcaagtcg
     3841 ctcatgttaa ggctgagaga gatatcctgg ctgaagctga caatgaatgg gtagttcgtc
     3901 tatattattc attccaagat aaggacaatt tatactttgt aatggactac attcctgggg
     3961 gtgatatgat gagcctatta attagaatgg gcatctttcc agaaagtctg gcacgattct
     4021 acatagcaga acttacctgt gcagttgaaa gtgttcataa aatgggtttt attcatagag
     4081 atattaaacc tgataatatt ttgattgatc gtgatggtca tattaaattg actgactttg
     4141 gcctctgcac tggcttcaga tggacacacg attctaagta ctatcagagt ggtgaccatc
     4201 cacggcaaga tagcatggat ttcagtaatg aatgggggga tccctcaagc tgtcgatgtg
     4261 gagacagact gaagccatta gagcggagag ctgcacgcca gcaccagcga tgtctagcac
     4321 attctttggt tgggactccc aattatattg cacctgaagt gttgctacga acaggataca
     4381 cacagttgtg tgattggtgg agtgttggtg ttattctttt tgaaatgttg gtgggacaac
     4441 ctcctttctt ggcacaaaca ccattagaaa cacaaatgaa ggttatcaac tggcaaacat
     4501 ctcttcacat tccaccacaa gctaaactca gtcctgaagc ttctgatctt attattaaac
     4561 tttgccgagg acccgaagat cgcttaggca agaatggtgc tgatgaaata aaagctcatc
     4621 cattttttaa aacaattgac ttctccagtg acctgagaca gcagtctgct tcatacattc
     4681 ctaaaatcac acacccaaca gatacatcaa attttgatcc tgttgatcct gataaattat
     4741 ggagtgatga taacgaggaa gaaaatgtaa atgacactct caatggatgg tataaaaatg
     4801 gaaagcatcc tgaacatgca ttctatgaat ttaccttccg aaggtttttt gatgacaatg
     4861 gctacccata taattatccg aagcctattg aatatgaata cattaattca caaggctcag
     4921 agcagcagtc ggatgaagat gatcaaaaca caggctcaga gattaaaaat cgcgatctag
     4981 tatatgttgt ttaaacgaat tcgggctcgg tacccgggga tcctctagag tcgacctgca
     5041 ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     5101 accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag
     5161 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5221 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     5281 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     5341 aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta
     5401 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     5461 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     5521 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5701 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5761 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5821 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5881 aagcttaatt aactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     5941 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     6001 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6061 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6121 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     6181 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6241 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6301 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6361 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6421 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6481 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     6541 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6601 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6661 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6721 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6781 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata
     6841 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc
     6901 cgctcatgcg atgataagct gtcaaacatg agaattacaa cttatatcgt atggggctga
     6961 cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat tttatctgat
     7021 taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg
     7081 aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc
     7141 gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac
     7201 cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt
     7261 gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7321 tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa
     7381 ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa
     7441 ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct
     7501 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
     7561 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt
     7621 atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc
     7681 agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac
     7741 atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga
     7801 caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct gatgtccggc
     7861 ggtgcttttg ccgttacgca ccaccccgtc atgagattac gcgttttagc ggtgcgtaca
     7921 attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc
     7981 agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
