LOCUS       sequence-479816-        6488 bp    DNA     circular SYN 07-AUG-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 6488)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6488)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 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            "Unpublished"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..6488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2368..2386)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2369..2386)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2506..2809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2964..2984
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3017..3064
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3096..3890
                     /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(3150..3169)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3760..3779
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4107..4652
                     /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     4553..4572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4769..4786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4936..4984
                     /note="synthetic polyadenylation signal"
     CDS             complement(5008..6045)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5021..5029
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(6076..6433)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6090..6225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6242..6433
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6390..6410
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaacagagc aagccacata gctggggggc agaggctcag
      841 cggctgggat aaggctgttc tcataagggt tggagtaggg gccatcggat aagccccctt
      901 gggctccctg ggagtcccct gagctgtagt cagttgagat gccagagtca gataccacaa
      961 ggtacaggta ctttaggtgg ggtggggtag ggggcagctc agggcacagt gtccatggac
     1021 gcaagagctg ggagctgggg tccaggatag tgtactcaaa gctggcagca gaggctccct
     1081 ctgggctggg cttcgaggcc aaagcagatg agcaggagga tgcttctgag ccttcatcca
     1141 tggccactat gtccacactg ccaccaggcc ctgggaggtc ctcactgggc gggttccggg
     1201 gcagcaacca tttgtccagc accagatagg tatcctgggc atgctcactg cccactggct
     1261 ccagcagggg gccctcatca tctgtccccg gctccactgc ctgcatcgtc ccccagcagc
     1321 gctctgagag gacttccagg gaagcaggtg ggtcctccgt gaagggggtg caggggctcc
     1381 accacaggca gccatcattc tggtacagcc acagctggaa gttacccttg tgggtggtga
     1441 agaggccttc aaactcgctc tctgggctcg ggatgccagg ccagatcttc tgcttcagag
     1501 cccggcggtg ggagagcagc gcgagcacgg tcagcagcac caggatgacc acgaggatga
     1561 gggagagcgt caggatgagg gggtccaggt cgctaggcgt cagcagcgac acaggctccg
     1621 accaggcgct ccagaagccg ccgaagctcg gctcagccat acgcgcgcgg acggcgaagg
     1681 tgtagcgcgt ccggccccgc aggttgctca gcacacactc ggtgcggccc tccaggatct
     1741 ccaccctctg tacgctccct gcgccgttgc cggccgagac gtccacctcg tagcggatgt
     1801 gagacgtcat gggtgtctca ggcggcggga gccagcgcaa cactacgtgg ccgctctcgt
     1861 cagccaaccg cgccaccagc cccacggggg cgtctaggag cactacttca ttgatgtgga
     1921 tgacacggtg atatcgcgga gcgccggagg ctgctgtgac gcgcaactct aggggcacga
     1981 agctcgacgt gtcggctgta ggcagcgaac accagaagcg caccgcacca cgagccgtgg
     2041 gagcctggtg caggcgacac agcttccatg gctcatcctc gagctggtag gagaagctgt
     2101 agttgcccgg gcccacccca gcgctcgccg cttcctccca gaaacacacc aagtcctcca
     2161 accgctcggt gaagcacaga agctcttcgg gcccccgggc cgccagcaag gccgctttgc
     2221 tctcgaactt ggggtccggg aggttaggcg ggggcgccca ggcggcccca gcgagcagga
     2281 gacaaaggga gccgacctgg ggccagaggg acgccccgag gtggtccatg gcgatcgcta
     2341 gctgcaaaaa gaacaagtag cttgtattct atagtgtcac ctaaatgggc gatctgacgg
     2401 ttcactaaac cagctctgct tatataggcc tccctggcgc gccactagtt attaatagta
     2461 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2521 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2581 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2641 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     2701 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     2761 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2821 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2881 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2941 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcaa
     3001 atcgaattct aatactgttg acaattaatc atcggcatag tatatcggca tagtataata
     3061 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     3121 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3181 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3241 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3301 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3361 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3421 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3481 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3541 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3601 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3661 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3721 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3781 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3841 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3901 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtcaaacatg agaattacaa cttatatcgt
     4021 atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat
     4081 tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     4141 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     4201 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     4261 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     4321 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     4381 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     4441 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     4501 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     4561 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt
     4621 gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc
     4681 cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc
     4741 cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat
     4801 cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc
     4861 acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct
     4921 gatgtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     4981 tattgcggcc atcgtgatgg ctagagtcta aactcgacct acctccttag cgcgcggtcg
     5041 tgtactgggc cggcggttgc cgctgtcggc cagcacctcg acgcagccgt cggtccatac
     5101 ggctgcgctc cggcgagcga tttgtgtgcg accgacggtg ccggccccgc tgcggacgat
     5161 ggcatcgcag cggccttgag cccaggcagc atcgtcgaag ttgccgtcca cgaggctctg
     5221 gtagagctga tccaggccga tgcgcagcat gtaggctcgc agacgagggc tgccggccag
     5281 ctcgggatgc cggcgctcga agtagcgagt ctgctgctcc atgcaagcca gccagggccg
     5341 ccagaagaag atgttggcca cctcgtactg actgtccccg aacatagctt cggaccagtc
     5401 gattacggcg gtgatgcggc cgttgtcggt caggacgttg ttgctgccga agtcggcatg
     5461 gacgaggtgg cgcacctcgg gacagtcttc ggcccacagc atcagttcgt ccagggcttg
     5521 agctacgctg gcggacacgg tgtcgtccat cacggtctgc cagtggtaga catggggatc
     5581 agcaatggcg caaatgaaat cccgccaggt tgtgtactgg ccgatgcctt ggggcccgaa
     5641 tgggccgaag ccgctggttt gactgaggtc ggcggcggca atagcatcca tagcttcggc
     5701 gacaggctgt aacacagcag gcagctctgt ttcggggagg tcttggagag taacgccttg
     5761 ggcgcgtcta ctgatgcagt atgtcaggct ctcgctgaac tcgccgatgt ccaacacttc
     5821 ggggatgggt agtgcagcgc tggcgaagtg gcggtacacg tagcggtctt tgtagaagcc
     5881 atcagcgcag ctattcaccc gcagtacata gccgcgtccg ccgacatcga agctgaaggc
     5941 tcggctctct tcgccctccg acaactgcat caggtcgctc acactgtcga acttctcgat
     6001 gagaaatttt tcaacgctgg tagcggtgag ttcgggcttc ttcatggtgg cgctagtgtc
     6061 agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     6121 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     6181 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     6241 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     6301 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6361 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     6421 ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg
     6481 tgctgcgc
//
