LOCUS       sequence-472399-        5533 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express NanoLuc(R)-HSD17B13 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 5533)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5533)
  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: NanoLuc-HSD17B13 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5533
                     /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(2612..2630)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2667..2788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2678..2697)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2732..2751
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3033..3390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3033..3224
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3056..3076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3241..3376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3405..3452
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3484..4278
                     /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(3538..3557)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4148..4167
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4342..4390
                     /note="synthetic polyadenylation signal"
     rep_origin      4547..5092
                     /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     4993..5012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5209..5226
                     /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 gaacattatt cttgaaatcc
     1621 ttctccttct gatcactatc atatatagtt accttgaaag tttggtaaaa tttttcattc
     1681 ctcaacggcg gaagtctgtg gccggtgaga tagtcctgat tactggcgca ggacatggaa
     1741 tcggccgcca gacgacgtac gagtttgcta agcgacaatc catactggta ctctgggata
     1801 ttaacaagcg gggtgtagaa gagacagcag ccgagtgtcg caagctgggc gtgactgcac
     1861 acgcatatgt cgtggactgc agtaatcggg aggagattta ccgatccctt aaccaggtaa
     1921 agaaggaggt aggagatgta acaatagttg taaataatgc cggaacggtc tatcctgcgg
     1981 atcttctcag tacaaaggat gaagaaatca caaaaacttt tgaagtgaat atactggggc
     2041 acttctggat aacaaaggcc ctccttccca gtatgatgga acggaaccat ggccacatcg
     2101 taaccgttgc gtccgtgtgc ggacatgagg gcataccata tttgattccc tattgcagca
     2161 gtaagtttgc ggccgtcggt tttcacaggg gattgacaag tgaactgcaa gcgttgggaa
     2221 agacgggtat taaaacctcc tgcctttgtc ccgtgtttgt caacacaggc tttacgaaga
     2281 accccagcac acgattgtgg ccggtgctcg agaccgatga ggtagtgcgg agccttatag
     2341 atggtatcct caccaacaaa aaaatgatct ttgttccctc atatatcaat atttttctgc
     2401 gattgcaaaa atttctgcct gagcgggcta gtgcgatact gaaccggatg caaaacatac
     2461 aattcgaggc ggtcgtcggt cacaagataa agatgaaagt ttaaacgaat tcgggctcgg
     2521 tacccgggga tcctctagag tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa
     2581 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2641 ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2701 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2761 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2821 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     2881 aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     2941 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     3001 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     3061 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3121 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3181 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3241 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3301 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3361 ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac aattaatcat
     3421 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag
     3481 atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3541 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3601 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3661 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3721 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3781 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3841 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3901 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3961 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4021 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4081 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4141 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4201 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4261 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4321 acctgccatc acgatggccg caataaaata tctttatttt cattacatct gtgtgttggt
     4381 tttttgtgtg aatcgatagc gataaggatc cgctcatgcg atgataagct gtcaaacatg
     4441 agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
     4501 gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg
     4561 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     4621 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     4681 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     4741 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     4801 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     4861 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     4921 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     4981 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     5041 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     5101 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg
     5161 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
     5221 agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt
     5281 tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca
     5341 gtatacactc cgctagcgct gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc
     5401 atgagattac gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg
     5461 tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag
     5521 aggatcgaga tct
//
