LOCUS       sequence-475067-        5924 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express HiBiT-GPR75 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 5924)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HiBiT-GPR75 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5924
                     /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"
     primer_bind     complement(2867..2885)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2922..3043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2933..2952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2987..3006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3288..3645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3288..3479
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3311..3331)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3496..3631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3660..3707
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3739..4533
                     /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(3793..3812)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4403..4422
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4597..4645
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4884..5472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4964..4983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5588..5871
                     /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 caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgaact
     1141 caacaggcca ccttcaggat gcccccaatg ccacctcgct ccatgtgcct cactcacagg
     1201 aaggaaacag cacctctctc caggagggtc ttcaggatct catccacaca gccaccttgg
     1261 tgacctgtac ttttctactg gcggtcatct tctgcctggg ttcctatggc aacttcattg
     1321 tcttcttgtc cttcttcgat ccagccttca ggaaattcag aaccaacttt gatttcatga
     1381 tcctgaacct gtccttctgt gacctcttca tttgtggagt gacagccccc atgttcacct
     1441 ttgtgttatt cttcagctca gccagtagta tcccggatgc tttctgcttc actttccatc
     1501 tcaccagttc aggcttcatc atcatgtctc tgaagacagt ggcagtgatc gccctgcacc
     1561 ggctccggat ggtgttgggg aaacagccta atcgcacggc ctcctttccc tgcaccgtac
     1621 tcctcaccct gcttctctgg gccaccagtt tcacccttgc caccttggct accttgaaaa
     1681 ccagcaagtc ccacctctgt cttcccatgt ccagtctgat tgctggaaaa gggaaagcca
     1741 ttttgtctct ctatgtggtc gacttcacct tctgtgttgc tgtggtctct gtctcttaca
     1801 tcatgattgc tcagaccctg cggaagaacg ctcaagtcag aaagtgcccc cctgtaatca
     1861 cagtcgatgc ttccagacca cagcctttca tgggggtccc tgtgcaggga ggtggagatc
     1921 ccatccagtg tgccatgccg gctctgtata ggaaccagaa ttacaacaaa ctgcagcacg
     1981 ttcagacccg tggatatacc aagagtccca accaactggt cacccctgca gcaagccgac
     2041 tccagctcgt atcagccatc aacctctcca ctgccaagga ttccaaagcc gtggtcacct
     2101 gtgtgatcat tgtgctgtca gtcctggtgt gctgtcttcc actggggatt tccttggtac
     2161 aggtggttct ctccagcaat gggagcttca ttctttacca gtttgaattg tttggattta
     2221 ctcttatatt tttcaagtca ggattaaacc cttttatata ttctcggaac agtgcagggc
     2281 tgagaaggaa agtgctctgg tgcctccaat acataggcct gggttttttc tgctgcaaac
     2341 aaaagactcg acttcgagcc atgggaaaag ggaacctcga agtcaacaga aacaaatcct
     2401 cccatcatga aacaaactct gcctacatgt tatctccaaa gccacagaag aaatttgtgg
     2461 accaggcttg tggcccaagt cattcaaaag aaagtatggt gagtcccaag atctctgctg
     2521 gacatcaaca ctgtggtcag agcagctcga cccccatcaa cactcggatt gaaccttact
     2581 acagcatcta taacagcagc ccttcccagg aggagagcag cccatgtaac ttacagccag
     2641 taaactcttt tggatttgcc aattcatata ttgccatgca ttatcacacc actaatgact
     2701 tagtgcagga atatgacagc acttcagcca agcagattcc agtcccctcc gttgtttaaa
     2761 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     2821 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2881 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2941 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3001 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3061 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     3121 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     3181 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     3241 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     3301 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3481 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3541 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3601 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg
     3661 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3721 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3781 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3841 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3901 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3961 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4021 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4081 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4141 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4201 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4261 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4321 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4381 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4441 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4501 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4561 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     4621 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg
     4681 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     4741 ggactggctt tctacccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4801 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4861 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4921 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4981 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5041 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5101 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5161 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5221 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5281 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5341 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5401 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5461 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5521 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5581 ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca
     5641 tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc
     5701 ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt
     5761 gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac
     5821 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     5881 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
