LOCUS       sequence-472462-        6165 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express CCDC6-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 6165)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6165)
  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: CCDC6-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..6165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(863..882)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(864..882)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(924..943)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          926..1058
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(1194..1397)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1223..1243)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1398..1777
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1968..2251
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      2367..2955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2856..2875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3194..3242
                     /note="synthetic polyadenylation signal"
     CDS             complement(3306..4100)
                     /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(3417..3436)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4027..4046
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4132..4179)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4194..4551)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4208..4343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4360..4551
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4508..4528
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4796..4917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4833..4852)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4887..4906
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4954..4972
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5060..5572)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
ORIGIN
        1 aatgctgtaa ctgagcagct ctcagttgct tcttcagccg ttccacttca ttctttaaaa
       61 acctgatgtg acgcatcata ttttctggag aatcaatctc catggagata tctctaggcg
      121 atggtggagc agagacgggc tggtctaatt tttcctgcag gattcgcttt tcagcttcaa
      181 gcttatccat ccttttccag aggcgattaa ctagtgcttc ttgttcttgt tccaatgtat
      241 tttcaaggtc aatcttctcc cgtctcaact gttctaatgt aagttgctta gaaatggtgt
      301 cattctccag ttttttaatt ttcttcatca gtttgttgac ctgaaattcc tgctcttgtt
      361 caagatgctg ttctagttcg gctttctcat gctgcaactg catcaatttt ctggagagct
      421 cattagtgag gaattcttct tctttctcat aatttacagc aagggtttct ttctccttct
      481 gcaaagcctg aattttcttg aataaagtgt tactaatgaa ttcttcttcc tgctcagccc
      541 tggcttggat ggtcacgctg gctttgcgca ggtcgcggtt ctcctcctgc agtgccttgc
      601 acttcagttt gtaggtctcc agctctatct tcagcacctt gttctcttgc tgcagcgagg
      661 ccaggcggtt ggtgagctcc tccaggcgga acggcgagat gacaatgccc cccgacttcc
      721 caccgccgcc gcctcccccg ccgccaccgc cgccgcccga ggtcgacgag caggacgact
      781 gcatggcggc cgagctgctg ctgttgcccc ccgccccgtc cgtgtcgctc tcgctggcgc
      841 tgtccgccat ggcgatcgct agccctatag tgagtcgtat taatactcta gccttaagag
      901 ctgtaattga actgggagtg gacacctgtg gagagaaagg caaagtggat gtcagtaaga
      961 ccaataggtg cctatcagaa acgcaagagt cttctctgtc tcgacaagcc cagtttctat
     1021 tggtctcctt aaacctgtct tgtaaccttg atacttacct gcccagtgcc tcacgaccaa
     1081 cttctgcagc ttaagttcga gactgttgtg tcagaagcac tgactgcgtt agcaatttaa
     1141 ctgtgataaa ctaccgcaat aaagcttcta gtgatctgac ggttcactaa accagctctg
     1201 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaacgg ggcggggtta
     1261 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     1321 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attggtgtac
     1381 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     1441 ggaaagtccc gtaaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     1501 acgtcaatag ggggcggact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     1561 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     1621 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     1681 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     1741 attgattact attaataact agtcaataat caatgccaac atggcggtca tattggacat
     1801 gagccaatat aaatgtacat attatgatat agatacaacg tatgcaatgg ccaatagcca
     1861 atattgattt atgctatata accaatgaat aatatggcta atggccaata ttgaagatct
     1921 cgatcctcta cgccggacgc atcgtggccg gagtactgcg gtttatctcg atggctacga
     1981 gggcagacag taagtggatt taccataatc ccttaattgt acgcaccgct aaaacgcgtt
     2041 cagcgcgatc acggcagcag acaggtaaaa atggcaacaa accacccgaa aaactgccgc
     2101 aatcgcgcct gataaatttt aaccgtatga atacctatgc aaccagaggg tacaggccac
     2161 attaccccca cttaatccac tgaagctgcc atttttcatg gtttcaccat cccagcgaag
     2221 ggccatccag cgtgcgttcc tgtatttccg gactataaaa ggatctaggt gaagatcctt
     2281 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2341 cccgtagaaa agatcaaagg atcttcttga aatccttttt ttctgcgcgt aatctgctgc
     2401 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2461 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2521 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2581 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2641 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2701 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2761 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2821 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2881 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2941 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3001 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3061 gcctttgagt gagctgatac cgggtagaaa gccagtccgc agaaacggtg ctgaccccgg
     3121 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gaggatcctt
     3181 atcgctatcg attcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     3241 ttgcggccat cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     3301 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     3361 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     3421 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     3481 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     3541 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     3601 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     3661 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     3721 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     3781 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     3841 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     3901 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     3961 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     4021 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     4081 aatccatctt gttcaatcat gatctgcctc ctgggtttag ttcctcacct tgtcgtatta
     4141 tactatgccg atatactatg ccgatgatta attgtcaaca gttaattaag ctttttgcaa
     4201 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     4261 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     4321 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     4381 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     4441 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     4501 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct
     4561 aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgcagat ccgcgtatgc
     4621 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     4681 taatattttg ttaaaattcg attttaccac atttgtagag gttttacttg cttaaaaaaa
     4741 cctcccacat ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4801 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4861 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4921 tgtctgctcg aagcggccgc cccaaggggt tatgctagtt attgctcagc ggtggcagca
     4981 gccaactcag cttcctttcg ggctttgtta gcagccggat cagcttgcat gcctgcaggt
     5041 cgactctaga gtcgcggcct tacgccagaa tgcgttcgca cagccgccag ccggtcactc
     5101 cgttgatggt tactcggaac agcagggagc cgtcggggtt gatcaggcgc tcgtcgataa
     5161 ttttgttgcc gttccacagg gtccctgtta cagtgatctt tttgccgtcg aacacggcga
     5221 tgccttcata cggccgtccg aaatagtcga tcatgttcgg cgtaaccccg tcgattacca
     5281 gtgtgccata gtgcaggatc accttaaagt gatgatcatc cacagggtac accaccttaa
     5341 aaattttttc gatctggccc atttggtcgc cgctcagacc ttcatacggg atgatgacat
     5401 ggatgtcgat cttcagccca ttttcaccgc tcaggacaat cctttggatc ggagttacgg
     5461 acaccccgag attctgaaac aaactggaca cacctccctg ttcaaggact tggtccaggt
     5521 tgtagccggc tgtctgtcgc cagtccccaa cgaaatcttc gagtgtgaag acgccgctcg
     5581 agccgagaga aacaggctgg gaggaggggt gcgccgaatg ttgcgaagga gtaggctgcg
     5641 aggtggctgc tgaggggacc gtgggctgca tgggtggcgg aggtggaggc ggaggtggct
     5701 ggactggggt ctgtgtgttg ggagatggag gcggcgtggg ccgtttgaat ttgtcaggac
     5761 tgttgcttct ccgtggtgaa ggccttgtga taccatggga tgttcccatg tgctgcacgt
     5821 gaagacccgg ggaattgtaa taagacattc cagctctagt cagtgaagtt ggtggcgtga
     5881 aaccaaccgt gtgacttgca tatgatagac caggtgatat aggcctgctt gaactcggag
     5941 aaggtgtgta agggatcggg ctggacacag tgcgaggtct taatccttgt gcagacatct
     6001 cattaaaata cctttcgtcg tccatttcta agctggactc actctcggag agctgtcgac
     6061 agagggcttc tcttctctcc atctccctct gcagcttcct ctggagcctc aagttctctt
     6121 ctctcatgtg acgttcctcc tccagatact gtgccatttt ctctg
//
