LOCUS       sequence-475128-        6227 bp    DNA     circular SYN 16-JUN-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)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6227)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: GPR75-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..6227
                     /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             2706..3218
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3306..3324)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3361..3482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3372..3391)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3426..3445
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3727..4084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3727..3918
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3750..3770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3935..4070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4099..4146
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4178..4972
                     /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(4232..4251)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4842..4861
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5036..5084
                     /note="synthetic polyadenylation signal"
     rep_origin      5241..5786
                     /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     5687..5706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5903..5920
                     /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 cgccatgaac tcaacaggcc
     1081 accttcagga tgcccccaat gccacctcgc tccatgtgcc tcactcacag gaaggaaaca
     1141 gcacctctct ccaggagggt cttcaggatc tcatccacac agccaccttg gtgacctgta
     1201 cttttctact ggcggtcatc ttctgcctgg gttcctatgg caacttcatt gtcttcttgt
     1261 ccttcttcga tccagccttc aggaaattca gaaccaactt tgatttcatg atcctgaacc
     1321 tgtccttctg tgacctcttc atttgtggag tgacagcccc catgttcacc tttgtgttat
     1381 tcttcagctc agccagtagt atcccggatg ctttctgctt cactttccat ctcaccagtt
     1441 caggcttcat catcatgtct ctgaagacag tggcagtgat cgccctgcac cggctccgga
     1501 tggtgttggg gaaacagcct aatcgcacgg cctcctttcc ctgcaccgta ctcctcaccc
     1561 tgcttctctg ggccaccagt ttcacccttg ccaccttggc taccttgaaa accagcaagt
     1621 cccacctctg tcttcccatg tccagtctga ttgctggaaa agggaaagcc attttgtctc
     1681 tctatgtggt cgacttcacc ttctgtgttg ctgtggtctc tgtctcttac atcatgattg
     1741 ctcagaccct gcggaagaac gctcaagtca gaaagtgccc ccctgtaatc acagtcgatg
     1801 cttccagacc acagcctttc atgggggtcc ctgtgcaggg aggtggagat cccatccagt
     1861 gtgccatgcc ggctctgtat aggaaccaga attacaacaa actgcagcac gttcagaccc
     1921 gtggatatac caagagtccc aaccaactgg tcacccctgc agcaagccga ctccagctcg
     1981 tatcagccat caacctctcc actgccaagg attccaaagc cgtggtcacc tgtgtgatca
     2041 ttgtgctgtc agtcctggtg tgctgtcttc cactggggat ttccttggta caggtggttc
     2101 tctccagcaa tgggagcttc attctttacc agtttgaatt gtttggattt actcttatat
     2161 ttttcaagtc aggattaaac ccttttatat attctcggaa cagtgcaggg ctgagaagga
     2221 aagtgctctg gtgcctccaa tacataggcc tgggtttttt ctgctgcaaa caaaagactc
     2281 gacttcgagc catgggaaaa gggaacctcg aagtcaacag aaacaaatcc tcccatcatg
     2341 aaacaaactc tgcctacatg ttatctccaa agccacagaa gaaatttgtg gaccaggctt
     2401 gtggcccaag tcattcaaaa gaaagtatgg tgagtcccaa gatctctgct ggacatcaac
     2461 actgtggtca gagcagctcg acccccatca acactcggat tgaaccttac tacagcatct
     2521 ataacagcag cccttcccag gaggagagca gcccatgtaa cttacagcca gtaaactctt
     2581 ttggatttgc caattcatat attgccatgc attatcacac cactaatgac ttagtgcagg
     2641 aatatgacag cacttcagcc aagcagattc cagtcccctc cgttgtttct ctcggctcga
     2701 gcggcgtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
     2761 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
     2821 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     2881 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     2941 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     3001 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     3061 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     3121 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     3181 tgaccggctg gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agagtcgacc
     3241 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     3301 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     3361 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3421 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3481 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     3541 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     3601 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3661 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     3721 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4021 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4081 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     4141 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     4201 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4261 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4321 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4381 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4441 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4501 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4561 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4621 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4681 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4741 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4801 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4861 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4921 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4981 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     5041 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     5101 gatccgctca tgcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg
     5161 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     5221 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     5281 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     5341 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     5401 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     5461 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5521 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     5581 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     5641 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     5701 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     5761 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     5821 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     5881 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     5941 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     6001 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     6061 cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga ttacgcgttt tagcggtgcg
     6121 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     6181 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
