LOCUS       sequence-477702-        6294 bp    DNA     circular SYN 02-JUL-2025
DEFINITION  Express HER3(1-664)-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 6294)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6294)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 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, HER3(1-664)-SmBiT
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3357..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3368..3387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3593..3626)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3636..3683
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3715..4509
                     /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(3769..3788)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4379..4398
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4667..5255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4747..4766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5381..5429
                     /note="synthetic polyadenylation signal"
     CDS             complement(5453..5851)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5848..5857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5882..6239)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5896..6031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6048..6239
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6196..6216
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat gagggcgaac
     1201 gacgctctgc aggtgctggg cttgcttttc agcctggccc ggggctccga ggtgggcaac
     1261 tctcaggcag tgtgtcctgg gactctgaat ggcctgagtg tgaccggcga tgctgagaac
     1321 caataccaga cactgtacaa gctctacgag aggtgtgagg tggtgatggg gaaccttgag
     1381 attgtgctca cgggacacaa tgccgacctc tccttcctgc agtggattcg agaagtgaca
     1441 ggctatgtcc tcgtggccat gaatgaattc tctactctac cattgcccaa cctccgcgtg
     1501 gtgcgaggga cccaggtcta cgatgggaag tttgccatct tcgtcatgtt gaactataac
     1561 accaactcca gccacgctct gcgccagctc cgcttgactc agctcaccga gattctgtca
     1621 gggggtgttt atattgagaa gaacgataag ctttgtcaca tggacacaat tgactggagg
     1681 gacatcgtga gggaccgaga tgctgagata gtggtgaagg acaatggcag aagctgtccc
     1741 ccctgtcatg aggtttgcaa ggggcgatgc tggggtcctg gatcagaaga ctgccagaca
     1801 ttgaccaaga ccatctgtgc tcctcagtgt aatggtcact gctttgggcc caaccccaac
     1861 cagtgctgcc atgatgagtg tgccgggggc tgctcaggcc ctcaggacac agactgcttt
     1921 gcctgccggc acttcaatga cagtggagcc tgtgtacctc gctgtccaca gcctcttgtc
     1981 tacaacaagc taactttcca gctggaaccc aatccccaca ccaagtatca gtatggagga
     2041 gtttgtgtag ccagctgtcc ccataacttt gtggtggatc aaacatcctg tgtcagggcc
     2101 tgtcctcctg acaagatgga agtagataaa aatgggctca agatgtgtga gccttgtggg
     2161 ggactatgtc ccaaagcctg tgagggaaca ggctctggga gccgcttcca gactgtggac
     2221 tcgagcaaca ttgatggatt tgtgaactgc accaagatcc tgggcaacct ggactttctg
     2281 atcaccggcc tcaatggaga cccctggcac aagatccctg ccctggaccc agagaagctc
     2341 aatgtcttcc ggacagtacg ggagatcaca ggttacctga acatccagtc ctggccgccc
     2401 cacatgcaca acttcagtgt tttttccaat ttgacaacca ttggaggcag aagcctctac
     2461 aaccggggct tctcattgtt gatcatgaag aacttgaatg tcacatctct gggcttccga
     2521 tccctgaagg aaattagtgc tgggcgtatc tatataagtg ccaataggca gctctgctac
     2581 caccactctt tgaactggac caaggtgctt cgggggccta cggaagagcg actagacatc
     2641 aagcataatc ggccgcgcag agactgcgtg gcagagggca aagtgtgtga cccactgtgc
     2701 tcctctgggg gatgctgggg cccaggccct ggtcagtgct tgtcctgtcg aaattatagc
     2761 cgaggaggtg tctgtgtgac ccactgcaac tttctgaatg gggagcctcg agaatttgcc
     2821 catgaggccg aatgcttctc ctgccacccg gaatgccaac ccatggaggg cactgccaca
     2881 tgcaatggct cgggctctga tacttgtgct caatgtgccc attttcgaga tgggccccac
     2941 tgtgtgagca gctgccccca tggagtccta ggtgccaagg gcccaatcta caagtaccca
     3001 gatgttcaga atgaatgtcg gccctgccat gagaactgca cccaggggtg taaaggacca
     3061 gagcttcaag actgtttagg acaaacactg gtgctgatcg gcaaaaccca tctgacaatg
     3121 gctttgacag tgatagcagg attggtagtg attttcatga tgctgggcgg cacttttctc
     3181 gtttctcaag gcagttcagg tggtggcggg agcggaggtg gaggctcgag cggtgtgacc
     3241 ggctaccggc tgttcgagga gattctgtaa ggccgcgact ctagagtcga cctgcaggca
     3301 tgcaagctga tttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag
     3361 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3421 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3481 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     3541 aagcaagtaa aacctctaca aatgtggtaa aatcttaatt aagaattcta atataacttc
     3601 gtatagcata cattatacga agttatggat ccactgttga caattaatca tcggcatagt
     3661 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     3721 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3781 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3841 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3901 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3961 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4021 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4081 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4141 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4201 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4261 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4321 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4381 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4441 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4501 ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5101 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5221 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5341 aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac
     5401 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga gtttagccct
     5461 cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac
     5521 tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc
     5581 cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc
     5641 cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa
     5701 aatgatatac attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga
     5761 cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg
     5821 tggattcttc ttgagacaaa ggcttggcca tggtggcgct agtgtcagaa gaatcgagct
     5881 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     5941 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     6001 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     6061 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     6121 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     6181 ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga
     6241 aggtacctaa ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgc
//
