LOCUS       sequence-486827-        6227 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Express GPR75-NanoLuc 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 6227)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6227)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: GPR75-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..6227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..79
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(167..679)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2333..2352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2334..2352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2394..2413)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2396..2528
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2664..2867)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2693..2713)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2868..3247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3692..3709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3826..4371)
                     /direction=LEFT
                     /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     complement(3906..3925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4528..4576
                     /note="synthetic polyadenylation signal"
     CDS             complement(4640..5434)
                     /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(4751..4770)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5361..5380
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5466..5513)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5528..5885)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5542..5677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5694..5885
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5842..5862
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    join(6130..6227,1..24)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6167..6186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     join(6221..6227,1..13)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccgcccc aaggggttat
       61 gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc tttgttagca
      121 gccggatcag cttgcatgcc tgcaggtcga ctctagagtc gcggccttac gccagaatgc
      181 gttcgcacag ccgccagccg gtcactccgt tgatggttac tcggaacagc agggagccgt
      241 cggggttgat caggcgctcg tcgataattt tgttgccgtt ccacagggtc cctgttacag
      301 tgatcttttt gccgtcgaac acggcgatgc cttcatacgg ccgtccgaaa tagtcgatca
      361 tgttcggcgt aaccccgtcg attaccagtg tgccatagtg caggatcacc ttaaagtgat
      421 gatcatccac agggtacacc accttaaaaa ttttttcgat ctggcccatt tggtcgccgc
      481 tcagaccttc atacgggatg atgacatgga tgtcgatctt cagcccattt tcaccgctca
      541 ggacaatcct ttggatcgga gttacggaca ccccgagatt ctgaaacaaa ctggacacac
      601 ctccctgttc aaggacttgg tccaggttgt agccggctgt ctgtcgccag tccccaacga
      661 aatcttcgag tgtgaagacg ccgctcgagc cgagagaaac aacggagggg actggaatct
      721 gcttggctga agtgctgtca tattcctgca ctaagtcatt agtggtgtga taatgcatgg
      781 caatatatga attggcaaat ccaaaagagt ttactggctg taagttacat gggctgctct
      841 cctcctggga agggctgctg ttatagatgc tgtagtaagg ttcaatccga gtgttgatgg
      901 gggtcgagct gctctgacca cagtgttgat gtccagcaga gatcttggga ctcaccatac
      961 tttcttttga atgacttggg ccacaagcct ggtccacaaa tttcttctgt ggctttggag
     1021 ataacatgta ggcagagttt gtttcatgat gggaggattt gtttctgttg acttcgaggt
     1081 tcccttttcc catggctcga agtcgagtct tttgtttgca gcagaaaaaa cccaggccta
     1141 tgtattggag gcaccagagc actttccttc tcagccctgc actgttccga gaatatataa
     1201 aagggtttaa tcctgacttg aaaaatataa gagtaaatcc aaacaattca aactggtaaa
     1261 gaatgaagct cccattgctg gagagaacca cctgtaccaa ggaaatcccc agtggaagac
     1321 agcacaccag gactgacagc acaatgatca cacaggtgac cacggctttg gaatccttgg
     1381 cagtggagag gttgatggct gatacgagct ggagtcggct tgctgcaggg gtgaccagtt
     1441 ggttgggact cttggtatat ccacgggtct gaacgtgctg cagtttgttg taattctggt
     1501 tcctatacag agccggcatg gcacactgga tgggatctcc acctccctgc acagggaccc
     1561 ccatgaaagg ctgtggtctg gaagcatcga ctgtgattac aggggggcac tttctgactt
     1621 gagcgttctt ccgcagggtc tgagcaatca tgatgtaaga gacagagacc acagcaacac
     1681 agaaggtgaa gtcgaccaca tagagagaca aaatggcttt cccttttcca gcaatcagac
     1741 tggacatggg aagacagagg tgggacttgc tggttttcaa ggtagccaag gtggcaaggg
     1801 tgaaactggt ggcccagaga agcagggtga ggagtacggt gcagggaaag gaggccgtgc
     1861 gattaggctg tttccccaac accatccgga gccggtgcag ggcgatcact gccactgtct
     1921 tcagagacat gatgatgaag cctgaactgg tgagatggaa agtgaagcag aaagcatccg
     1981 ggatactact ggctgagctg aagaataaca caaaggtgaa catgggggct gtcactccac
     2041 aaatgaagag gtcacagaag gacaggttca ggatcatgaa atcaaagttg gttctgaatt
     2101 tcctgaaggc tggatcgaag aaggacaaga agacaatgaa gttgccatag gaacccaggc
     2161 agaagatgac cgccagtaga aaagtacagg tcaccaaggt ggctgtgtgg atgagatcct
     2221 gaagaccctc ctggagagag gtgctgtttc cttcctgtga gtgaggcaca tggagcgagg
     2281 tggcattggg ggcatcctga aggtggcctg ttgagttcat ggcgatcgct agccctatag
     2341 tgagtcgtat taatactcta gccttaagag ctgtaattga actgggagtg gacacctgtg
     2401 gagagaaagg caaagtggat gtcagtaaga ccaataggtg cctatcagaa acgcaagagt
     2461 cttctctgtc tcgacaagcc cagtttctat tggtctcctt aaacctgtct tgtaaccttg
     2521 atacttacct gcccagtgcc tcacgaccaa cttctgcagc ttaagttcga gactgttgtg
     2581 tcagaagcac tgactgcgtt agcaatttaa ctgtgataaa ctaccgcaat aaagcttcta
     2641 gtgatctgac ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta
     2701 ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc ccgttgattt
     2761 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     2821 tcaaaccgct atccacgccc attggtgtac tgccaaaacc gcatcaccat ggtaatagcg
     2881 atgactaata cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat gtactgggca
     2941 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcggact tggcatatga
     3001 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     3061 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     3121 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     3181 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     3241 caatgccaac atggcggtca tattggacat gagccaatat aaatgtacat attatgatat
     3301 agatacaacg tatgcaatgg ccaatagcca atattgattt atgctatata accaatgaat
     3361 aatatggcta atggccaata ttgaagatct cgatcctcta cgccggacgc atcgtggccg
     3421 gagtactgcg gtttatctcg atggctacga gggcagacag taagtggatt taccataatc
     3481 ccttaattgt acgcaccgct aaaacgcgta atctcatgac ggggtggtgc gtaacggcaa
     3541 aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat
     3601 gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc
     3661 agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
     3721 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
     3781 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
     3841 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     3901 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
     3961 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
     4021 cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
     4081 agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg
     4141 caaaagcacc actggcagca gccactggta attgatttag aggagttagt cttgaagtca
     4201 tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag
     4261 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     4321 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     4381 cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca
     4441 cctgaagtca gccccatacg atataagttg taattctcat gtttgacagc ttatcatcgc
     4501 atgagcggat ccttatcgct atcgattcac acaaaaaacc aacacacaga tgtaatgaaa
     4561 ataaagatat tttattgcgg ccatcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt
     4621 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     4681 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     4741 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     4801 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     4861 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     4921 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     4981 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     5041 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     5101 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     5161 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     5221 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     5281 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     5341 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     5401 gagaacctgc gtgcaatcca tcttgttcaa tcatgatctg cctcctgggt ttagttcctc
     5461 accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc aacagttaat
     5521 taagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5581 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5641 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     5701 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5761 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5821 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     5881 ctcagaaggt acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
     5941 agatccgcgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6001 gaaattgtaa gcgttaatat tttgttaaaa ttcgatttta ccacatttgt agaggtttta
     6061 cttgcttaaa aaaacctccc acatctcccc ctgaacctga aacataaaat gaatgcaatt
     6121 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6181 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggttt
//
