LOCUS       sequence-485900-        8322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-AR 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 8322)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8322)
  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            "Unpublished"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: CMV-LgBiT-AR Expression Vector
FEATURES             Location/Qualifiers
     source          1..8322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        487..505
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     487..504
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    4034..4115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4071..4090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4132..5310
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4363..5301
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5258..5278
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5688..6482
                     /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(5742..5761)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6352..6371
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6506..6554
                     /note="synthetic polyadenylation signal"
     rep_origin      6680..7268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7169..7188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7440..8099)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8014..8033
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(8131..8178)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(8211..8231)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tggaaatccc cgtgagtcaa accgctatcc acgcccattg gtgtactgcc aaaaccgcat
       61 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccgtaa
      121 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
      181 cggacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
      241 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
      301 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
      361 tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg attactatta
      421 ataactagtg gcgcgccagg gaggcctata taagcagagc tggtttagtg aaccgtcaga
      481 tcgcccattt aggtgacact atagaataca agctacttgt tctttttgca gctagcgctc
      541 accatggtct tcacactcga agatttcgtt ggggactggg aacagacagc cgcctacaac
      601 ctggaccaag tccttgaaca gggaggtgtg tccagtttgc tgcagaatct cgccgtgtcc
      661 gtaactccga tccaaaggat tgtccggagc ggtgaaaatg ccctgaagat cgacatccat
      721 gtcatcatcc cgtatgaagg tctgagcgcc gaccaaatgg cccagatcga agaggtgttt
      781 aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcccta tggcacactg
      841 gtaatcgacg gggttacgcc gaacatgctg aactatttcg gacggccgta tgaaggcatc
      901 gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt
      961 atcgacgagc gcctgatcac ccccgacggc tccatgctgt tccgagtaac catcaacagt
     1021 gggagctccg gtggtggcgg gagcggaggt ggaggctcga gcggtgcgat cgccatggaa
     1081 gtgcagttag ggctgggaag ggtctaccct cggccgccgt ccaagaccta ccgaggagct
     1141 ttccagaatc tgttccagag cgtgcgcgaa gtgatccaga acccgggccc caggcaccca
     1201 gaggccgcga gcgcagcacc tcccggcgcc agtttgctgc tgctgcagca gcagcagcag
     1261 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcaaga gactagcccc
     1321 aggcagcagc agcagcagca gggtgaggat ggttctcccc aagcccatcg tagaggcccc
     1381 acaggctacc tggtcctgga tgaggaacag caaccttcac agccgcagtc ggccctggag
     1441 tgccaccccg agagaggttg cgtcccagag cctggagccg ccgtggccgc cagcaagggg
     1501 ctgccgcagc agctgccagc acctccggac gaggatgact cagctgcccc atccacgttg
     1561 tccctgctgg gccccacttt ccccggctta agcagctgct ccgctgacct taaagacatc
     1621 ctgagcgagg ccagcaccat gcaactcctt cagcaacagc agcaggaagc agtatccgaa
     1681 ggcagcagca gcgggagagc gagggaagcc tcgggggctc ccacttcctc caaggacaat
     1741 tacttagggg gcacttcgac catttctgac aacgccaagg agttgtgtaa ggcagtgtcg
     1801 gtgtccatgg gcctgggtgt ggaggcgttg gagcatctga gtccagggga acagcttcgg
     1861 ggggattgca tgtacgcccc acttttggga gttccacccg ctgtgcgtcc cactccttgt
     1921 gccccattgg ccgaatgcaa aggttctctg ctagacgaca gcgcaggcaa gagcactgaa
     1981 gatactgctg agtattcccc tttcaaggga ggttacacca aagggctaga aggcgagagc
     2041 ctaggctgct ctggcagcgc tgcagcaggg agctccggga cacttgaact gccgtctacc
     2101 ctgtctctct acaagtccgg agcactggac gaggcagctg cgtaccagag tcgcgactac
     2161 tacaactttc cactggctct ggccggaccg ccgccccctc cgccgcctcc ccatccccac
     2221 gctcgcatca agctggagaa cccgctggac tacggcagcg cctgggcggc tgcggcggcg
     2281 cagtgccgct atggggacct ggcgagcctg catggcgcgg gtgcagcggg acccggttct
     2341 gggtcaccct cagccgccgc ttcctcatcc tggcacactc tcttcacagc cgaagaaggc
     2401 cagttgtatg gaccgtgtgg tggtggtggg ggtggtggcg gcggcggcgg cggcggcggc
     2461 ggcggcggcg gcggcggcgg cggcggcggc gaggcgggag ctgtagcccc ctacggctac
     2521 actcggcccc ctcaggggct ggcgggccag gaaagcgact tcaccgcacc tgatgtgtgg
     2581 taccctggcg gcatggtgag cagagtgccc tatcccagtc ccacttgtgt caaaagcgaa
     2641 atgggcccct ggatggatag ctactccgga ccttacgggg acatgcgttt ggagactgcc
     2701 agggaccatg ttttgcccat tgactattac tttccacccc agaagacctg cctgatctgt
     2761 ggagatgaag cttctgggtg tcactatgga gctctcacat gtggaagctg caaggtcttc
     2821 ttcaaaagag ccgctgaagg gaaacagaag tacctgtgcg ccagcagaaa tgattgcact
     2881 attgataaat tccgaaggaa aaattgtcca tcttgtcgtc ttcggaaatg ttatgaagca
     2941 gggatgactc tgggagcccg gaagctgaag aaacttggta atctgaaact acaggaggaa
     3001 ggagaggctt ccagcaccac cagccccact gaggagacaa cccagaagct gacagtgtca
     3061 cacattgaag gctatgaatg tcagcccatc tttctgaatg tcctggaagc cattgagcca
     3121 ggtgtagtgt gtgctggaca cgacaacaac cagcccgact cctttgcagc cttgctctct
     3181 agcctcaatg aactgggaga gagacagctt gtacacgtgg tcaagtgggc caaggccttg
     3241 cctggcttcc gcaacttaca cgtggacgac cagatggctg tcattcagta ctcctggatg
     3301 gggctcatgg tgtttgccat gggctggcga tccttcacca atgtcaactc caggatgctc
     3361 tacttcgccc ctgatctggt tttcaatgag taccgcatgc acaagtcccg gatgtacagc
     3421 cagtgtgtcc gaatgaggca cctctctcaa gagtttggat ggctccaaat caccccccag
     3481 gaattcctgt gcatgaaagc actgctactc ttcagcatta ttccagtgga tgggctgaaa
     3541 aatcaaaaat tctttgatga acttcgaatg aactacatca aggaactcga tcgtatcatt
     3601 gcatgcaaaa gaaaaaatcc cacatcctgc tcaagacgct tctaccagct caccaagctc
     3661 ctggactccg tgcagcctat tgcgagagag ctgcatcagt tcacttttga cctgctaatc
     3721 aagtcacaca tggtgagcgt ggactttccg gaaatgatgg cagagatcat ctctgtgcaa
     3781 gtgcccaaga tcctttctgg gaaagtcaag cccatctatt tccacaccca ggtttaaacg
     3841 aattcgggct cggtacccgg ggatcctcta gaaacaccta ggctctcgta cgaagtcagc
     3901 tgataacgtc gacgtattgc ggccgcggat cataatcagc cataccacat ttgtagaggt
     3961 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4021 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4081 cacaaatttc acaaataaag catttttttc actgccgacg tcacgcgtct tggctccggt
     4141 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     4201 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     4261 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     4321 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     4381 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     4441 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     4501 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     4561 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     4621 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     4681 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     4741 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     4801 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     4861 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     4921 gcggaaagat ggccgcttcc cggccctgct gcaggcagct caaaatggag gacgcggcgc
     4981 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     5041 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     5101 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     5161 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     5221 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     5281 agtttttttc ttccatttca ggtgtcgtga aagccgaatt ctgcagatat ccatcacact
     5341 gtcgagcatg catctagtcc agtgtggtgg aattcggctt cgtggggacg ccgtacaggg
     5401 acgtgcacct ctcccgctgc accgcctcca gcgtcgccgc cggctcgaag gacggggccg
     5461 ggatgacgat gcaggcggcg tgggaggtgg cgcccaagtt gcccatgacc atgccgaagc
     5521 agtggtagaa gggcaccggc agacacaccc ggtcctgctc cgtgtagccg accgtgcggc
     5581 ccacccagta gccgttgttg aggatgttgt ggtgggagag cgtggcgccc ttggggaagc
     5641 cggtgtgccg gaggtgtact ggatgttgac cgggaatcga tgccaccatg attgaacaag
     5701 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5761 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5821 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5881 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5941 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     6001 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     6061 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     6121 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     6181 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     6241 tcgtgaccca cggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     6301 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6361 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6421 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6481 gaactctagc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     6541 tggttttttg tgtgacgatt ccggactata aaaggatcta ggtgaagatc ctttttgata
     6601 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6661 aaaagatcaa aggatcttct tgaaatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7321 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     7381 agtgagctga taccggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     7441 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     7501 tggaagccat cacagacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     7561 ccttgcgtat aatatttgcc catagtgaaa acgggggcga agaagttgtc catattggcc
     7621 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     7681 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     7741 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     7801 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     7861 ccgtctttca ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga
     7921 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     7981 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     8041 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatg
     8101 atctgcctcc tgggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc
     8161 cgatgattaa ttgtcaacag tattagaatt cgatttgacc tcccaccgta cacgcctacc
     8221 gcccatttgc gtcaacgggg cggggttatt acgacatttt ggaaagtccc gttgattttg
     8281 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga ct
//
