LOCUS       sequence-470652-        5384 bp    DNA     circular SYN 02-MAY-2025
DEFINITION  Express LgBiT-BAK 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 5384)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5384)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBiT Assay Vector, BAK
FEATURES             Location/Qualifiers
     source          1..5384
                     /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(2327..2345)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2382..2503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2393..2412)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2447..2466
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2748..3105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2748..2939
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2771..2791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2956..3091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3120..3167
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3199..3993
                     /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(3253..3272)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3863..3882
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4057..4105
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4344..4932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4424..4443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5048..5331
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg gaacagacag ccgcctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg ctgcagaatc tcgccgtgtc cgtaactccg atccaaagga
     1201 ttgtccggag cggtgaaaat gccctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgc cgaccaaatg gcccagatcg aagaggtgtt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgccct atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgct gaactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 cccccgacgg ctccatgctg ttccgagtaa ccatcaacag cggaggaggt ggctcaggtg
     1561 gagggggctc cgcgatcgcc atggcttcgg ggcaaggccc aggtcctccc aggcaggagt
     1621 gcggagagcc tgccctgccc tctgcttctg aggagcaggt agcccaggac acagaggagg
     1681 ttttccgcag ctacgttttt taccgccatc agcaggaaca ggaggctgaa ggggtggctg
     1741 cccctgccga cccagagatg gtcaccttac ctctgcaacc tagcagcacc atggggcagg
     1801 tgggacggca gctcgccatc atcggggacg acatcaaccg acgctatgac tcagagttcc
     1861 agaccatgtt gcagcacctg cagcccacgg cagagaatgc ctatgagtac ttcaccaaga
     1921 ttgccaccag cctgtttgag agtggcatca attggggccg tgtggtggct cttctgggct
     1981 tcggctaccg tctggcccta cacgtctacc agcatggcct gactggcttc ctaggccagg
     2041 tgacccgctt cgtggtcgac ttcatgctgc atcactgcat tgcccggtgg attgcacaga
     2101 ggggtggctg ggtggcagcc ctgaacttgg gcaatggtcc catcctgaac gtgctggtgg
     2161 ttctgggtgt ggttctgttg ggccagtttg tggtacgaag attcttcaaa tcagtttaaa
     2221 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     2281 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2341 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2401 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2461 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2521 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2581 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     2641 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     2701 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     2761 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg
     3121 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3181 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3241 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3301 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3361 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3421 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3481 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3541 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3601 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3661 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3721 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3781 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3841 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3901 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3961 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4021 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     4081 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg
     4141 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     4201 ggactggctt tctacccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4741 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4921 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5041 ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat ggtgaaacca
     5101 tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc
     5161 ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt cgggtggttt
     5221 gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac
     5281 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     5341 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
