LOCUS       sequence-470781-        8986 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express NanoLuc(R)-FANCD2 (WT) 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 8986)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8986)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-FANCD2 (WT) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8986
                     /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(6065..6083)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6120..6241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6131..6150)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6185..6204
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6486..6843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6486..6677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6509..6529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6694..6829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6858..6905
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6937..7731
                     /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(6991..7010)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7601..7620
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7795..7843
                     /note="synthetic polyadenylation signal"
     rep_origin      8000..8545
                     /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     8446..8465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8662..8679
                     /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 ggtttccaaa agaagactgt
     1621 caaaatctga ggataaagag agcctgacag aagatgcctc caaaaccagg aagcaaccac
     1681 tttccaaaaa gacaaagaaa tctcatattg ctaatgaagt tgaagaaaat gacagcatct
     1741 ttgtaaagct tcttaagata tcaggaatta ttcttaaaac gggagagagt cagaatcaac
     1801 tagctgtgga tcaaatagct ttccaaaaga agctctttca gaccctgagg agacaccctt
     1861 cctatcccaa aataatagaa gaatttgtta gtggcctgga gtcttacatt gaggatgaag
     1921 acagtttcag gaactgcctt ttgtcttgtg agcgtctgca ggatgaggaa gccagtatgg
     1981 gtgcatctta ttctaagagt ctcatcaaac tgcttctggg gattgacata ctgcagcctg
     2041 ccattatcaa aaccttattt gagaagttgc cagaatattt ttttgaaaac aagaacagtg
     2101 atgaaatcaa catacctcga ctcattgtca gtcaactaaa atggcttgac agagttgtgg
     2161 atggcaagga cctcaccacc aagatcatgc agctgatcag tattgctcca gagaacctgc
     2221 agcatgacat catcaccagc ctacctgaga tcctagggga ttcccagcac gctgatgtgg
     2281 ggaaagaact cagtgaccta ctgatagaga atacttcact cactgtccca atcctggatg
     2341 tcctttcaag cctccgactt gacccaaact tcctattgaa ggttcgccag ttggtgatgg
     2401 ataagttgtc gtctattaga ttggaggatt tacctgtgat aataaagttc attcttcatt
     2461 ccgtaacagc catggataca cttgaggtaa tttctgagct tcgggagaag ttggatctgc
     2521 agcattgtgt tttgccatca cggttacagg cttcccaagt aaagttgaaa agtaaaggac
     2581 gagcaagttc ctcaggaaat caagaaagca gcggtcagag ctgtattatt ctcctctttg
     2641 atgtaataaa gtcagctatt agatatgaga aaaccatttc agaagcctgg attaaggcaa
     2701 ttgaaaacac tgcctcagta tctgaacaca aggtgtttga cctggtgatg cttttcatca
     2761 tctatagcac caatactcag acaaagaagt acattgacag ggtgctaaga aataagattc
     2821 gatcaggctg cattcaagaa cagctgctcc agagtacatt ctctgttcat tacttagttc
     2881 ttaaggatat gtgttcatcc attctgtcgc tggctcagag tttgcttcac tctctagacc
     2941 agagtataat ttcatttggc agtctcctat acaaatatgc atttaagttt tttgacacgt
     3001 actgccagca ggaagtggtt ggtgccttag tgacccatat ctgcagtggg aatgaagctg
     3061 aagttgatac tgccttagat gtccttctag agttggtagt gttaaaccca tctgctatga
     3121 tgatgaatgc tgtctttgta aagggcattt tagattatct ggataacata tcccctcagc
     3181 aaatacgaaa actcttctat gttctcagca cactggcatt tagcaaacag aatgaagcca
     3241 gcagccacat ccaggatgac atgcacttgg tgataagaaa gcagctctct agcaccgtat
     3301 tcaagtacaa gctcattggg attattggtg ctgtgaccat ggctggcatc atggcggcag
     3361 acagaagtga atcacctagt ttgacccaag agagagccaa cctgagcgat gagcagtgca
     3421 cacaggtgac ctccttgttg cagttggttc attcctgcag tgagcagtct cctcaggcct
     3481 ctgcacttta ctatgatgaa tttgccaacc tgatccaaca tgaaaagctg gatccaaaag
     3541 ccctggaatg ggttgggcat accatctgta atgatttcca ggatgccttc gtagtggact
     3601 cctgtgttgt tccggaaggt gactttccat ttcctgtgaa agcactgtac ggactggaag
     3661 aatacgacac tcaggatggg attgccataa acctcctgcc gctgctgttt tctcaggact
     3721 ttgcaaaaga tgggggtccg gtgacctcac aggaatcagg ccaaaaattg gtgtctccgc
     3781 tgtgcctggc tccgtatttc cggttactga gactttgtgt ggagagacag cataacggaa
     3841 acttggagga gattgatggt ctactagatt gtcctatatt cctaactgac ctggagcctg
     3901 gagagaagtt ggagtccatg tctgctaaag agcgttcatt catgtgttct ctcatatttc
     3961 ttactctcaa ctggttccga gagattgtaa atgccttctg ccaggaaaca tcacctgaga
     4021 tgaaggggaa ggtgctcact cggttaaagc acattgtaga attgcaaata atcctggaaa
     4081 agtacttggc agtcacccca gactatgtcc ctcctcttgg aaactttgat gtggaaactt
     4141 tagatataac acctcatact gttactgcta tttcagcaaa aatcagaaag aaaggaaaaa
     4201 tagaaaggaa acaaaaaaca gatggcagca agacatcctc ctctgacaca ctttcagaag
     4261 agaaaaattc agaatgtgac cctacgccat ctcatagagg ccagctaaac aaggagttca
     4321 cagggaagga agaaaagaca tcattgttac tacataattc ccatgctttt ttccgagagc
     4381 tggacattga ggtcttctct attctacatt gtggacttgt gacgaagttc atcttagata
     4441 ctgaaatgca cactgaagct acagaagttg tgcaacttgg gccccctgag ctgcttttct
     4501 tgctggaaga tctctcccag aagctggaga gtatgctgac acctcctatt gccaggagag
     4561 tcccctttct caagaacaaa ggaagccgga atattggatt ctcacatctc caacagagat
     4621 ctgcccaaga aattgttcat tgtgtttttc aactgctgac cccaatgtgt aaccacctgg
     4681 agaacattca caactatttt cagtgtttag ctgctgagaa tcacggtgta gttgatggac
     4741 caggagtgaa agttcaggag taccacataa tgtcttcctg ctatcagagg ctgctgcaga
     4801 tttttcatgg gctttttgct tggagtggat tttctcaacc tgaaaatcag aatttactgt
     4861 attcagccct ccatgtcctt agtagccgac tgaaacaggg agaacacagc cagcctttgg
     4921 aggaactact cagccagagc gtccattact tgcagaattt ccatcaaagc attcccagtt
     4981 tccagtgtgc tctttatctc atcagacttt tgatggttat tttggagaaa tcaacagctt
     5041 ctgctcagaa caaagaaaaa attgcttccc ttgccagaca attcctctgt cgggtgtggc
     5101 caagtgggga taaagagaag agcaacatct ctaatgacca gctccatgct ctgctctgta
     5161 tctacctgga gcacacagag agcattctga aggccataga ggagattgct ggtgttggtg
     5221 tcccagaact gatcaactct cctaaagatg catcttcctc cacattccct acactgacca
     5281 ggcatacttt tgttgttttc ttccgtgtga tgatggctga actagagaag acggtgaaaa
     5341 aaattgagcc tggcacagca gcagactcgc agcagattca tgaagagaaa ctcctctact
     5401 ggaacatggc tgttcgagac ttcagtatcc tcatcaactt gataaaggta tttgatagtc
     5461 atcctgttct gcatgtatgt ttgaagtatg ggcgtctctt tgtggaagca tttctgaagc
     5521 aatgtatgcc gctcctagac ttcagtttta gaaaacaccg ggaagatgtt ctgagcttac
     5581 tggaaacctt ccagttggac acaaggctgc ttcatcacct gtgtgggcat tccaagattc
     5641 accaggacac gagactcacc caacatgtgc ctctgctcaa aaagaccctg gaacttttag
     5701 tttgcagagt caaagctatg ctcactctca acaattgtag agaggctttc tggctgggca
     5761 atctaaaaaa ccgggacttg cagggtgaag agattaagtc ccaaaattcc caggagagca
     5821 cagcagatga gagtgaggat gacatgtcat cccaggcctc caagagcaaa gccactgagg
     5881 atggtgaaga agacgaagta agtgctggag aaaaggagca agatagtgat gagagttatg
     5941 atgactctga tgtttaaacg aattcgggct cggtacccgg ggatcctcta gagtcgacct
     6001 gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     6061 gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat
     6121 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     6181 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     6241 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     6301 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     6361 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     6421 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     6481 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     6541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     6601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6661 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6781 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6841 aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc atagtataat
     6901 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     6961 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     7021 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     7081 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     7141 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     7201 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     7261 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     7321 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7381 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7441 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7501 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7561 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7621 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7681 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7741 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     7801 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     7861 atccgctcat gcgatgataa gctgtcaaac atgagaatta caacttatat cgtatggggc
     7921 tgacttcagg tgctacattt gaagagataa attgcactga aatctagaaa tattttatct
     7981 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     8041 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     8101 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     8161 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     8221 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     8281 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     8341 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     8401 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     8461 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     8521 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     8581 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     8641 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat
     8701 cacatattct gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac
     8761 tgacaccctc atcagtgcca acatagtaag ccagtataca ctccgctagc gctgatgtcc
     8821 ggcggtgctt ttgccgttac gcaccacccc gtcatgagat tacgcgtttt agcggtgcgt
     8881 acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac
     8941 cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
