LOCUS       sequence-470761-        5567 bp    DNA     circular SYN 02-MAY-2025
DEFINITION  Express NanoLuc(R)-15-PGDH 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 5567)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5567)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-15-PGDH Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5567
                     /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(2510..2528)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2565..2686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2576..2595)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2630..2649
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2931..3288
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2931..3122
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2954..2974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3139..3274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3303..3350
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3382..4176
                     /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(3436..3455)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4046..4065
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4240..4288
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4527..5115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4607..4626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5231..5514
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 gcacgtgaac ggcaaagtgg
     1621 cgctggtgac cggcgcggct cagggcatag gcagagcctt tgcagaggcg ctgctgctta
     1681 agggcgccaa ggtagcgctg gtggattgga atcttgaagc aggtgtacag tgtaaagctg
     1741 ccctggatga gcagtttgaa cctcagaaga ctctgttcat ccagtgcgat gtggctgacc
     1801 agcaacaact gagagacact tttagaaaag ttgtagacca ctttggaaga ctggacattt
     1861 tggtcaataa tgctggagtg aataatgaga aaaactggga aaaaactctg caaattaatt
     1921 tggtttctgt tatcagtgga acctatcttg gtttggatta catgagtaag caaaatggag
     1981 gtgaaggcgg catcattatc aatatgtcat ctttagcagg actcatgccc gttgcacagc
     2041 agccggttta ttgtgcttca aagcatggca tagttggatt cacacgctca gcagcgttgg
     2101 ctgctaatct tatgaacagt ggtgtgagac tgaatgccat ttgtccaggc tttgttaaca
     2161 cagccatcct tgaatcaatt gaaaaagaag aaaacatggg acaatatata gaatataagg
     2221 atcatatcaa ggatatgatt aaatactatg gaattttgga cccaccattg attgccaatg
     2281 gattgataac actcattgaa gatgatgctt taaatggtgc tattatgaag atcacaactt
     2341 ctaagggaat tcattttcaa gactatgata caactccatt tcaagcaaaa acccaagttt
     2401 aaacgaattc gggctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     2461 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2521 taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga
     2581 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2641 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2701 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     2761 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     2821 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     2881 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     2941 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3061 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3121 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3181 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3241 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa
     3301 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3361 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3421 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3481 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3541 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3601 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3661 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3721 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3781 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3841 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3901 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3961 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4021 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4081 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4141 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4201 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     4261 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt
     4321 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     4381 tgcggactgg ctttctaccc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4441 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4501 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4561 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4621 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4681 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4741 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4801 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4861 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4921 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4981 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5041 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5101 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5161 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5221 cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa
     5281 ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt
     5341 tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg
     5401 tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg
     5461 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     5521 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
