LOCUS       sequence-471913-        5405 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express 15-PGDH-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 5405)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5405)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: 15-PGDH-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5405
                     /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             1884..2396
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2484..2502)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2539..2660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2550..2569)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2604..2623
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2905..3262
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2905..3096
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2928..2948)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3113..3248
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3277..3324
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3356..4150
                     /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(3410..3429)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4020..4039
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4214..4262
                     /note="synthetic polyadenylation signal"
     rep_origin      4419..4964
                     /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     4865..4884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5081..5098
                     /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 cgccatgcac gtgaacggca
     1081 aagtggcgct ggtgaccggc gcggctcagg gcataggcag agcctttgca gaggcgctgc
     1141 tgcttaaggg cgccaaggta gcgctggtgg attggaatct tgaagcaggt gtacagtgta
     1201 aagctgccct ggatgagcag tttgaacctc agaagactct gttcatccag tgcgatgtgg
     1261 ctgaccagca acaactgaga gacactttta gaaaagttgt agaccacttt ggaagactgg
     1321 acattttggt caataatgct ggagtgaata atgagaaaaa ctgggaaaaa actctgcaaa
     1381 ttaatttggt ttctgttatc agtggaacct atcttggttt ggattacatg agtaagcaaa
     1441 atggaggtga aggcggcatc attatcaata tgtcatcttt agcaggactc atgcccgttg
     1501 cacagcagcc ggtttattgt gcttcaaagc atggcatagt tggattcaca cgctcagcag
     1561 cgttggctgc taatcttatg aacagtggtg tgagactgaa tgccatttgt ccaggctttg
     1621 ttaacacagc catccttgaa tcaattgaaa aagaagaaaa catgggacaa tatatagaat
     1681 ataaggatca tatcaaggat atgattaaat actatggaat tttggaccca ccattgattg
     1741 ccaatggatt gataacactc attgaagatg atgctttaaa tggtgctatt atgaagatca
     1801 caacttctaa gggaattcat tttcaagact atgatacaac tccatttcaa gcaaaaaccc
     1861 aagtttctct cggctcgagc ggcgtcttca cactcgaaga tttcgttggg gactggcgac
     1921 agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc
     1981 agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt gaaaatgggc
     2041 tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac caaatgggcc
     2101 agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc
     2161 tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac tatttcggac
     2221 ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt
     2281 ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc
     2341 gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg gcgtaaggcc
     2401 gcgactctag agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa
     2461 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc
     2521 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2581 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2641 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2701 gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat
     2761 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     2821 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     2881 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     2941 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3001 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3061 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3121 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3181 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3241 tggaggccta ggcttttgca aaaagcttaa ttaactgttg acaattaatc atcggcatag
     3301 tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat
     3361 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3421 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3481 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3541 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3601 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3661 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3721 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3781 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3841 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3901 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3961 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4021 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4081 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4141 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4201 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     4261 tgaatcgata gcgataagga tccgctcatg cgatgataag ctgtcaaaca tgagaattac
     4321 aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa
     4381 atctagaaat attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc
     4441 gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc
     4501 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     4561 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     4621 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     4681 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     4741 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     4801 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     4861 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     4921 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
     4981 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
     5041 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
     5101 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
     5161 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
     5221 tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcatgagatt
     5281 acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct
     5341 cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga
     5401 gatct
//
