LOCUS       sequence-475276-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EPOR-LgBiT 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 6264)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: EPOR-LgBiT Vector
FEATURES             Location/Qualifiers
     source          1..6264
                     /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(3207..3225)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3262..3383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3273..3292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3327..3346
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3628..3985
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3628..3819
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3651..3671)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3836..3971
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4000..4047
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4079..4873
                     /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(4133..4152)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4743..4762
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4937..4985
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5224..5812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5304..5323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5928..6211
                     /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 ggctagcgat cgccatggac cacctcgggg
     1081 cgtccctctg gccccaggtc ggctcccttt gtctcctgct cgctggggcc gcctgggcgc
     1141 ccccgcctaa cctcccggac cccaagttcg agagcaaagc ggccttgctg gcggcccggg
     1201 ggcccgaaga gcttctgtgc ttcaccgagc ggttggagga cttggtgtgt ttctgggagg
     1261 aagcggcgag cgctggggtg ggcccgggca actacagctt ctcctaccag ctcgaggatg
     1321 agccatggaa gctgtgtcgc ctgcaccagg ctcccacggc tcgtggtgcg gtgcgcttct
     1381 ggtgttcgct gcctacagcc gacacgtcga gcttcgtgcc cctagagttg cgcgtcacag
     1441 cagcctccgg cgctccgcga tatcaccgtg tcatccacat caatgaagta gtgctcctag
     1501 acgcccccgt ggggctggtg gcgcggttgg ctgacgagag cggccacgta gtgttgcgct
     1561 ggctcccgcc gcctgagaca cccatgacgt ctcacatccg ctacgaggtg gacgtctcgg
     1621 ccggcaacgg cgcagggagc gtacagaggg tggagatcct ggagggccgc accgagtgtg
     1681 tgctgagcaa cctgcggggc cggacgcgct acaccttcgc cgtccgcgcg cgtatggctg
     1741 agccgagctt cggcggcttc tggagcgcct ggtcggagcc tgtgtcgctg ctgacgccta
     1801 gcgacctgga ccccctcatc ctgacgctct ccctcatcct cgtggtcatc ctggtgctgc
     1861 tgaccgtgct cgcgctgctc tcccaccgcc gggctctgaa gcagaagatc tggcctggca
     1921 tcccgagccc agagagcgag tttgaaggcc tcttcaccac ccacaagggt aacttccagc
     1981 tgtggctgta ccagaatgat ggctgcctgt ggtggagccc ctgcaccccc ttcacggagg
     2041 acccacctgc ttccctggaa gtcctctcag agcgctgctg ggggacgatg caggcagtgg
     2101 agccggggac agatgatgag ggccccctgc tggagccagt gggcagtgag catgcccagg
     2161 atacctatct ggtgctggac aaatggttgc tgccccggaa cccgcccagt gaggacctcc
     2221 cagggcctgg tggcagtgtg gacatagtgg ccatggatga aggctcagaa gcatcctcct
     2281 gctcatctgc tttggcctcg aagcccagcc cagagggagc ctctgctgcc agctttgagt
     2341 acactatcct ggaccccagc tcccagctct tgcgtccatg gacactgtgc cctgagctgc
     2401 cccctacccc accccaccta aagtacctgt accttgtggt atctgactct ggcatctcaa
     2461 ctgactacag ctcaggggac tcccagggag cccaaggggg cttatccgat ggcccctact
     2521 ccaaccctta tgagaacagc cttatcccag ccgctgagcc tctgcccccc agctatgtgg
     2581 cttgctctgt ttctcaaggc agttcaggtg gtggcgggag cggaggtgga ggctcgagcg
     2641 gtgtcttcac actcgaagat ttcgttgggg actgggaaca gacagccgcc tacaacctgg
     2701 accaagtcct tgaacaggga ggtgtgtcca gtttgctgca gaatctcgcc gtgtccgtaa
     2761 ctccgatcca aaggattgtc cggagcggtg aaaatgccct gaagatcgac atccatgtca
     2821 tcatcccgta tgaaggtctg agcgccgacc aaatggccca gatcgaagag gtgtttaagg
     2881 tggtgtaccc tgtggatgat catcacttta aggtgatcct gccctatggc acactggtaa
     2941 tcgacggggt tacgccgaac atgctgaact atttcggacg gccgtatgaa ggcatcgccg
     3001 tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg
     3061 acgagcgcct gatcaccccc gacggctcca tgctgttccg agtaaccatc aacagctaag
     3121 gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
     3181 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3241 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3301 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3361 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3421 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     3481 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3541 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     3601 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     3661 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3721 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3841 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3901 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3961 ttttggaggc ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca
     4021 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     4081 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4141 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4201 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4261 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4321 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4381 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4441 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4501 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4561 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4621 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4681 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4741 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4801 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4861 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4921 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4981 gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc
     5041 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt
     5101 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5161 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5221 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5281 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5341 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5401 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5461 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5521 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5581 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5641 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5701 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5761 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     5821 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5881 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     5941 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     6001 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     6061 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     6121 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     6181 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     6241 gtccggcgta gaggatcgag atct
//
