LOCUS       sequence-474684-        8975 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 8975)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8975)
  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..8975
                     /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(6054..6072)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6109..6230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6120..6139)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6174..6193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6475..6832
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6475..6666
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6498..6518)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6683..6818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6847..6894
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6926..7720
                     /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(6980..6999)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7590..7609
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7784..7832
                     /note="synthetic polyadenylation signal"
     rep_origin      7989..8534
                     /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     8435..8454
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8651..8668
                     /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 atagtgtgct
     4981 ctttatctca tcagactttt gatggttatt ttggagaaat caacagcttc tgctcagaac
     5041 aaagaaaaaa ttgcttccct tgccagacaa ttcctctgtc gggtgtggcc aagtggggat
     5101 aaagagaaga gcaacatctc taatgaccag ctccatgctc tgctctgtat ctacctggag
     5161 cacacagaga gcattctgaa ggccatagag gagattgctg gtgttggtgt cccagaactg
     5221 atcaactctc ctaaagatgc atcttcctcc acattcccta cactgaccag gcatactttt
     5281 gttgttttct tccgtgtgat gatggctgaa ctagagaaga cggtgaaaaa aattgagcct
     5341 ggcacagcag cagactcgca gcagattcat gaagagaaac tcctctactg gaacatggct
     5401 gttcgagact tcagtatcct catcaacttg ataaaggtat ttgatagtca tcctgttctg
     5461 catgtatgtt tgaagtatgg gcgtctcttt gtggaagcat ttctgaagca atgtatgccg
     5521 ctcctagact tcagttttag aaaacaccgg gaagatgttc tgagcttact ggaaaccttc
     5581 cagttggaca caaggctgct tcatcacctg tgtgggcatt ccaagattca ccaggacacg
     5641 agactcaccc aacatgtgcc tctgctcaaa aagaccctgg aacttttagt ttgcagagtc
     5701 aaagctatgc tcactctcaa caattgtaga gaggctttct ggctgggcaa tctaaaaaac
     5761 cgggacttgc agggtgaaga gattaagtcc caaaattccc aggagagcac agcagatgag
     5821 agtgaggatg acatgtcatc ccaggcctcc aagagcaaag ccactgagga tggtgaagaa
     5881 gacgaagtaa gtgctggaga aaaggagcaa gatagtgatg agagttatga tgactctgat
     5941 gtttaaacga attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca
     6001 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     6061 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     6121 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     6181 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     6241 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     6301 aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc
     6361 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     6421 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     6481 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6541 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     6601 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     6661 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6721 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6781 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa
     6841 ttaactgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     6901 aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc
     6961 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     7021 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     7081 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     7141 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7201 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7261 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7321 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7381 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7441 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     7501 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     7561 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7621 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7681 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     7741 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     7801 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgctcatg
     7861 cgatgataag ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt
     7921 gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga
     7981 tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc
     8041 gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     8101 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     8161 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     8221 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     8281 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     8341 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     8401 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     8461 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     8521 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     8581 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     8641 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     8701 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     8761 tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt
     8821 tgccgttacg caccaccccg tcatgagatt acgcgtttta gcggtgcgta caattaaggg
     8881 attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc
     8941 ggccacgatg cgtccggcgt agaggatcga gatct
//
