LOCUS       sequence-475275-        5823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EPOR-SmBiT 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 5823)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5823)
  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            "Unpublished"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-SmBiT Vector
FEATURES             Location/Qualifiers
     source          1..5823
                     /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(2766..2784)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2821..2942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2886..2905
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3187..3544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3187..3378
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3210..3230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3395..3530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3559..3606
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3638..4432
                     /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(3692..3711)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4302..4321
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4496..4544
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4783..5371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4863..4882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5487..5770
                     /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 gtgtgaccgg ctaccggctg ttcgaggaga ttctgtaagg ccgcgactct agagtcgacc
     2701 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2761 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     2821 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2881 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2941 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     3001 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     3061 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3121 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     3181 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3241 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3301 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3361 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3421 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3481 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3541 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3601 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     3661 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3721 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3781 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3841 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3901 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3961 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4021 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4081 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4141 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4201 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4261 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4321 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4381 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4441 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     4501 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     4561 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     4621 gggtcagcac cgtttctgcg gactggcttt ctacccggta tcagctcact caaaggcggt
     4681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5221 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5341 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     5401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5461 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     5521 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     5581 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     5641 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     5701 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     5761 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     5821 tct
//
