LOCUS       sequence-474694-        5302 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express LgBiT-MAFG 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 5302)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LgBiT-MAFG Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5302
                     /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(1356..1374)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1357..1374)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1494..1797
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1952..1972
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2005..2052
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2084..2878
                     /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(2138..2157)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2748..2767
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3036..3624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3116..3135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3750..3798
                     /note="synthetic polyadenylation signal"
     CDS             complement(3822..4859)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3835..3843
                     /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(4890..5247)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4904..5039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5056..5247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5204..5224
                     /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 ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 cgatcgggca tccgtcttgg actttactat tgtgatgacg ctggtggcgg ccaccttgcc
      361 tgggacgagg ggccccaggc cggcggcaag ggggccccgg gctggcgcca cggggctgcg
      421 ggccaccgtc cgggcgaagg tctgcagcgc ctcgtacttg gagcgcagcg cgtcgagctc
      481 cagcttcatg ctggcgttct ctgaggccag cttctccacc tcctgctgca gctccgcctt
      541 ctgcttctcc agctcctcct tctgcgtcac ccgcttcacg cggcagctgg cagcgtagcc
      601 gcggttcttg agcgtgcgcc ggcgctgctt cagctggacg atctcctcct tggacaggcc
      661 ccgcaggtgc tggttcagct cccgcaccga catggtcacc agctcctcat ccgtcaggct
      721 ggtgccattc tcacccggct cccgcttcac cttcaaggcc ttgtttcctt tattgggggt
      781 cgtcatggcg atcgcaccgc tcgagcctcc acctccgctc ccgccaccac cggagctccc
      841 actgttgatg gttactcgga acagcatgga gccgtcgggg gtgatcaggc gctcgtcgat
      901 aattttgttg ccgttccaca gggtccctgt tacagtgatc tttttgccgt cgaacacggc
      961 gatgccttca tacggccgtc cgaaatagtt cagcatgttc ggcgtaaccc cgtcgattac
     1021 cagtgtgcca tagggcagga tcaccttaaa gtgatgatca tccacagggt acaccacctt
     1081 aaacacctct tcgatctggg ccatttggtc ggcgctcaga ccttcatacg ggatgatgac
     1141 atggatgtcg atcttcaggg cattttcacc gctccggaca atcctttgga tcggagttac
     1201 ggacacggcg agattctgca gcaaactgga cacacctccc tgttcaagga cttggtccag
     1261 gttgtaggcg gctgtctgtt cccagtcccc aacgaaatct tcgagtgtga agaccatggt
     1321 gagcgctagc tgcaaaaaga acaagtagct tgtattctat agtgtcacct aaatgggcga
     1381 tctgacggtt cactaaacca gctctgctta tataggcctc cctggcgcgc cactagttat
     1441 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     1501 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     1561 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     1621 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
     1681 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     1741 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     1801 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
     1861 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     1921 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
     1981 gaggtcaaat cgaattctaa tactgttgac aattaatcat cggcatagta tatcggcata
     2041 gtataatacg acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg
     2101 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2161 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     2221 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     2281 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     2341 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     2401 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     2461 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     2521 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     2581 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     2641 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     2701 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     2761 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     2821 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt
     2881 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     2941 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3601 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     3661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3721 acctagatcc ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga
     3781 aaataaagat attttattgc ggccatcgtg atggctagag tctaaactcg acctacctcc
     3841 ttagcgcgcg gtcgtgtact gggccggcgg ttgccgctgt cggccagcac ctcgacgcag
     3901 ccgtcggtcc atacggctgc gctccggcga gcgatttgtg tgcgaccgac ggtgccggcc
     3961 ccgctgcgga cgatggcatc gcagcggcct tgagcccagg cagcatcgtc gaagttgccg
     4021 tccacgaggc tctggtagag ctgatccagg ccgatgcgca gcatgtaggc tcgcagacga
     4081 gggctgccgg ccagctcggg atgccggcgc tcgaagtagc gagtctgctg ctccatgcaa
     4141 gccagccagg gccgccagaa gaagatgttg gccacctcgt actgactgtc cccgaacata
     4201 gcttcggacc agtcgattac ggcggtgatg cggccgttgt cggtcaggac gttgttgctg
     4261 ccgaagtcgg catggacgag gtggcgcacc tcgggacagt cttcggccca cagcatcagt
     4321 tcgtccaggg cttgagctac gctggcggac acggtgtcgt ccatcacggt ctgccagtgg
     4381 tagacatggg gatcagcaat ggcgcaaatg aaatcccgcc aggttgtgta ctggccgatg
     4441 ccttggggcc cgaatgggcc gaagccgctg gtttgactga ggtcggcggc ggcaatagca
     4501 tccatagctt cggcgacagg ctgtaacaca gcaggcagct ctgtttcggg gaggtcttgg
     4561 agagtaacgc cttgggcgcg tctactgatg cagtatgtca ggctctcgct gaactcgccg
     4621 atgtccaaca cttcggggat gggtagtgca gcgctggcga agtggcggta cacgtagcgg
     4681 tctttgtaga agccatcagc gcagctattc acccgcagta catagccgcg tccgccgaca
     4741 tcgaagctga aggctcggct ctcttcgccc tccgacaact gcatcaggtc gctcacactg
     4801 tcgaacttct cgatgagaaa tttttcaacg ctggtagcgg tgagttcggg cttcttcatg
     4861 gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     4921 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     4981 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     5041 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     5101 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     5161 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     5221 ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt
     5281 atttcaggcc atggtgctgc gc
//
