LOCUS       sequence-472443-        8963 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express CDK13-NanoLuc(R) 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 8963)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 8963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CDK13-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8963
                     /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             3604..3615
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5442..5954
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(6042..6060)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6097..6218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6108..6127)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6162..6181
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6463..6820
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6463..6654
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6486..6506)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6671..6806
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6835..6882
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6914..7708
                     /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(6968..6987)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7578..7597
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7772..7820
                     /note="synthetic polyadenylation signal"
     rep_origin      7977..8522
                     /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     8423..8442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8639..8656
                     /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 ggctagcgat cgccatgccg agcagctcgg
     1081 acacggcgct ggggggaggc gggggcctga gctgggcgga gaagaagttg gaggaacgcc
     1141 gcaagcggag gcgattcctg tcccctcagc agccgccgct gctgttgccg ctcctgcagc
     1201 cgcagctcct gcaaccgccg ccgcccccgc cgcctctgct cttcctggct gctcccggca
     1261 cggccgccgc cgcagccgcc gccgccgcgg cctcctcctc ttgcttcagc ccgggccccc
     1321 ctctggaggt caagcggctg gcgagaggca agaggcgcgc aggagggcgg cagaagcggc
     1381 gtcgcgggcc ccgcgccggg caggaggcgg agaagcgtcg ggtcttctcg ctgccccagc
     1441 cgcagcagga cggcggtggc ggtgctagta gcggcggggg tgtgaccccg ctggtggaat
     1501 acgaggatgt gagctcccag tccgagcagg ggctgctgct ggggggggcc agcgcggcaa
     1561 cggcggcgac ggctgccggg ggaacggggg gcagcggcgg gagtccggcc tcctcctccg
     1621 gcacccmgcg gcgcggggrg gggkcggrgc gcaggccccg ccgggaccgc cgcagcagca
     1681 gtggccgcag caaggagcgc caccgcgagc accggcggcg ggatgggcag cgcggtggca
     1741 gcgaggcctc caagtcccgc agccgccaca gccacagcgg cgaggaacgg gccgaggtcg
     1801 ccaagagcgg cagcagcagc agcagcggcg gccgccggaa aagcgcttcg gccacatcca
     1861 gcagcagtag cagccgcaag gaccgggact cgaaggccca ccgcagccgg actaagtcgt
     1921 ccaaggagcc gccttcggcc tacaaggaac cgcccaaggc ctaccgggag gacaagaccg
     1981 agcctaaggc ctacaggcgg cggcggtccc tcagcccact gggaggccgg gacgacagcc
     2041 cggtgtccca cagggcctct cagagcctga ggagccgcaa gtcccccagc ccggcaggag
     2101 gtggcagcag cccctattct cggcggctgc cgcgctcccc gagcccctac agtcgccgcc
     2161 gctcccccag ctacagccgc cacagctcct acgagcgggg cggcgacgtg tcccctagtc
     2221 cctacagcag cagcagctgg cgccgctctc gcagtcccta cagccctgtg ctcagacggt
     2281 ctggaaaatc ccgaagcaga agcccgtatt catctaggca ttcaagatct cgtagcaggc
     2341 acagattgtc tagatccaga agtcgtcatt ctagtatttc tcctagcaca ctaactctga
     2401 agagtagcct ggcagctgaa ttgaacaaga ataaaaaagc acgagcagca gaggcagcaa
     2461 gagccgcaga agcagcgaaa gctgcagaag caactaaggc tgctgaggct gctgccaagg
     2521 ctgcaaaagc ttcaaacact tctacaccta ccaaggggaa cacggaaact agtgccagtg
     2581 catcacaaac aaaccatgtg aaggatgtga agaaaattaa aattgaacat gcaccttctc
     2641 cctcaagtgg tggaacttta aaaaatgaca aagcaaaaac aaagccacct cttcaggtaa
     2701 cgaaggtgga aaataatttg attgtagata aagccaccaa gaaagcagtc atagttggaa
     2761 aggagagtaa atctgctgct acaaaggagg aatcagtatc tcttaaagag aaaaccaaac
     2821 cacttacacc aagcatagga gccaaggaga aggagcaaca tgtagcttta gtcacctcta
     2881 cattaccacc gttacctttg cctcccatgc tgcctgaaga taaagaagct gatagcttac
     2941 gaggaaatat ttcagtaaaa gcagttaaaa aagaagtaga aaagaaactc cgatgtcttc
     3001 ttgctgattt accgctgccc cctgagctac caggaggaga tgatctttca aagagtccag
     3061 aggaaaagaa aacagcaaca cagttacata gtaaaaggag gcctaaaata tgtgggcctc
     3121 gctatggtga aaccaaagaa aaagatattg actggggaaa acgctgcgtg gataaatttg
     3181 atatcatcgg aattattgga gaaggtactt acggacaagt ttacaaagcc agggataaag
     3241 acactggaga aatggtagcc ttaaaaaaag tacgtctgga taatgaaaag gaaggctttc
     3301 caattacagc aattcgagaa attaaaattc tccggcagct tacccatcag agtattatca
     3361 atatgaagga aatagtgact gataaagaag atgctttgga tttcaagaag gacaaaggtg
     3421 cattttatct ggtgtttgaa tatatggacc atgatctgat gggactactg gaatcaggct
     3481 tggttcattt taatgaaaat cacataaagt catttatgag acagctcatg gagggtctgg
     3541 attattgtca taagaagaac tttttgcata gagatattaa atgttccaat atccttctaa
     3601 ataatagagg gcagataaaa cttgcagact ttggacttgc tcgattgtat agctcagaag
     3661 aaagtcggcc gtatactaac aaggtaatta ctttatggta ccgtccacct gaactgctac
     3721 tgggagaaga acgatacaca ccagccattg atgtatggag ctgtggctgt atccttggcg
     3781 aactcttcac taaaaaacct atatttcaag caaatcagga acttgcacaa ctagaattaa
     3841 taagccgaat atgtgggagt ccatgtcctg cagtgtggcc tgatgtaatc aaactaccat
     3901 atttcaacac catgaaacca aagaagcaat atcgtcgaaa gttaagagaa gaatttgttt
     3961 ttattcctgc agctgcgcta gacttatttg attacatgct tgccttggat cctagtaagc
     4021 gctgcactgc tgaacaggct cttcagtgcg agttcctccg agatgtggaa ccctcaaaaa
     4081 tgcctccacc agatctccct ttatggcaag attgtcatga gttatggagt aaaaagcgaa
     4141 gaagacagaa gcagatgggc atgactgatg atgtttccac aattaaagcc cccaggaagg
     4201 acttgtctct gggcttggat gacagcagaa ccaacacacc ccagggtgtg ctgccatctt
     4261 cacagctgaa atctcagggc agctcaaatg tggcacctgg tgaaaaacag acagatccat
     4321 caacaccaca acaggagtct tcgaaaccgt tgggaggaat tcagccttct tctcagacca
     4381 tccagcctaa agtggagact gatgctgccc aggcggctgt gcagagtgca tttgcagttc
     4441 tgttgactca gttaataaag gctcagcagt caaagcagaa agatgtgcta ctagaagaga
     4501 gggaaaatgg atcgggacat gaagcgtcat tacaactcag gccacctcca gaacctagca
     4561 ctccggtgtc gggacaagat gacctcatcc agcatcaaga tatgaggatc ttggagctaa
     4621 cgccagaacc agaccggcct cgaattctgc ctcctgacca acgacctccc gagcctcctg
     4681 aaccaccacc agtcactgag gaagatctag attatcggac agaaaaccag catgtaccca
     4741 ccaccagttc ttcattaact gaccctcatg ccggagtgaa ggcagccctg ttacagctgc
     4801 ttgctcagca tcagccccag gatgacccca aaagagaagg tgggattgat tatcaagcag
     4861 gagacactta cgtgtccact tcagactaca aggacaactt tggatcctct tctttctctt
     4921 ctgctcctta tgttagcaat gatggtctag gaagcagttc tgctccacca ctagaacgac
     4981 gtagtttcat tggaaattca gatattcagt ctttggataa ctacagtact gcttcatctc
     5041 attctggtgg tccacctcag ccttctgcct tttctgagtc atttcccagt tcagtagctg
     5101 gatatggaga catttacctc aatgctggtc ccatgttgtt tagtggagac aaggaccata
     5161 gatttgaata tagccatggt cctattgcag tcctggcaaa cagcagtgac ccttccacgg
     5221 ggccagagag tactcatcct ttgccagcaa agatgcacaa ctataactat ggtggtaact
     5281 tacaggaaaa tccgagtggc cccagcctca tgcatggaca gacctggact tctcctgccc
     5341 aaggacctgg atattcacaa ggatacaggg gacatattag cacatcaact ggcagaggca
     5401 gaggcagagg gttaccatac gtttctctcg gctcgagcgg cgtcttcaca ctcgaagatt
     5461 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
     5521 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
     5581 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     5641 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
     5701 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     5761 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     5821 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     5881 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     5941 gcattctggc gtaaggccgc gactctagag tcgacctgca ggcatgcaag ctgatccggc
     6001 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     6061 ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     6121 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6181 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     6241 ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca
     6301 aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     6361 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     6421 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     6481 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6541 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6601 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6661 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6721 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6781 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac
     6841 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     6901 caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6961 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     7021 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     7081 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     7141 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     7201 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     7261 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     7321 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     7381 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     7441 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     7501 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     7561 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     7621 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7681 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     7741 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     7801 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgctcatgcg atgataagct
     7861 gtcaaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     7921 gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga
     7981 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     8041 ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga
     8101 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     8161 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     8221 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     8281 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     8341 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     8401 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8461 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     8521 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     8581 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     8641 cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg
     8701 tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca
     8761 tagtaagcca gtatacactc cgctagcgct gatgtccggc ggtgcttttg ccgttacgca
     8821 ccaccccgtc atgagattac gcgttttagc ggtgcgtaca attaagggat tatggtaaat
     8881 ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg
     8941 tccggcgtag aggatcgaga tct
//
