LOCUS       sequence-486456-        6744 bp    DNA     circular SYN 19-SEP-2025
DEFINITION  Express HER1(1-668)-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 6744)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBiT Assay Vector, HER1(1-668)-LgBiT
FEATURES             Location/Qualifiers
     source          1..6744
                     /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(3687..3705)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3742..3863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3753..3772)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3807..3826
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4108..4465
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4108..4299
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4131..4151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4316..4451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4480..4527
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4559..5353
                     /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(4613..4632)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5223..5242
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5417..5465
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5704..6292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5784..5803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6408..6691
                     /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 ggctagcgat cgccatgcga ccctccggga
     1081 cggccggggc agcgctcctg gcgctgctgg ctgcgctctg cccggcgagt cgggctctgg
     1141 aggaaaagaa agtttgccaa ggcacgagta acaagctcac gcagttgggc acttttgaag
     1201 atcattttct cagcctccag aggatgttca ataactgtga ggtggtcctt gggaatttgg
     1261 aaattaccta tgtgcagagg aattatgatc tttccttctt aaagaccatc caggaggtgg
     1321 ctggttatgt cctcattgcc ctcaacacag tggagcgaat tcctttggaa aacctgcaga
     1381 tcatcagagg aaatatgtac tacgaaaatt cctatgcctt agcagtctta tctaactatg
     1441 atgcaaataa aaccggactg aaggagctgc ccatgagaaa tttacaggaa atcctgcatg
     1501 gcgccgtgcg gttcagcaac aaccctgccc tgtgcaacgt ggagagcatc cagtggcggg
     1561 acatagtcag cagtgacttt ctcagcaaca tgtcgatgga cttccagaac cacctgggca
     1621 gctgccaaaa gtgtgatcca agctgtccca atgggagctg ctggggtgca ggagaggaga
     1681 actgccagaa actgaccaaa atcatctgtg cccagcagtg ctccgggcgc tgccgtggca
     1741 agtcccccag tgactgctgc cacaaccagt gtgctgcagg ctgcacaggc ccccgggaga
     1801 gcgactgcct ggtctgccgc aaattccgag acgaagccac gtgcaaggac acctgccccc
     1861 cactcatgct ctacaacccc accacgtacc agatggatgt gaaccccgag ggcaaataca
     1921 gctttggtgc cacctgcgtg aagaagtgtc cccgtaatta tgtggtgaca gatcacggct
     1981 cgtgcgtccg agcctgtggg gccgacagct atgagatgga ggaagacggc gtccgcaagt
     2041 gtaagaagtg cgaagggcct tgccgcaaag tgtgtaacgg aataggtatt ggtgaattta
     2101 aagactcact ctccataaat gctacgaata ttaaacactt caaaaactgc acctccatca
     2161 gtggcgatct ccacatcctg ccggtggcat ttaggggtga ctccttcaca catactcctc
     2221 ctctggatcc acaggaactg gatattctga aaaccgtaaa ggaaatcaca gggtttttgc
     2281 tgattcaggc ttggcctgaa aacaggacgg acctccatgc ctttgagaac ctagaaatca
     2341 tacgcggcag gaccaagcaa catggtcagt tttctcttgc agtcgtcagc ctgaacataa
     2401 catccttggg attacgctcc ctcaaggaga taagtgatgg agatgtgata atttcaggaa
     2461 acaaaaattt gtgctatgca aatacaataa actggaaaaa actgtttggg acctccggtc
     2521 agaaaaccaa aattataagc aacagaggtg aaaacagctg caaggccaca ggccaggtct
     2581 gccatgcctt gtgctccccc gagggctgct ggggcccgga gcccaaggac tgcgtctctt
     2641 gccggaatgt cagccgaggc agggaatgcg tggacaagtg caaccttctg gagggtgagc
     2701 caagggagtt tgtggagaac tctgagtgca tacagtgcca cccagagtgc ctgcctcagg
     2761 ccatgaacat cacctgcaca ggacggggac cagacaactg tatccagtgt gcccactaca
     2821 ttgacggccc ccactgcgtc aagacctgcc cggcaggagt catgggagaa aacaacaccc
     2881 tggtctggaa gtacgcagac gccggccatg tgtgccacct gtgccatcca aactgcacct
     2941 acggatgcac agggccaggt cttgaaggct gtccaacgaa tgggcctaag atcccgtcca
     3001 tcgccactgg gatggtgggg gccctcctct tgctgctggt ggtggccctg gggatcggcc
     3061 tcttcatggt ttctcaaggc agttcaggtg gtggcgggag cggaggtgga ggctcgagcg
     3121 gtgtcttcac actcgaagat ttcgttgggg actgggaaca gacagccgcc tacaacctgg
     3181 accaagtcct tgaacaggga ggtgtgtcca gtttgctgca gaatctcgcc gtgtccgtaa
     3241 ctccgatcca aaggattgtc cggagcggtg aaaatgccct gaagatcgac atccatgtca
     3301 tcatcccgta tgaaggtctg agcgccgacc aaatggccca gatcgaagag gtgtttaagg
     3361 tggtgtaccc tgtggatgat catcacttta aggtgatcct gccctatggc acactggtaa
     3421 tcgacggggt tacgccgaac atgctgaact atttcggacg gccgtatgaa ggcatcgccg
     3481 tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg
     3541 acgagcgcct gatcaccccc gacggctcca tgctgttccg agtaaccatc aacagctaag
     3601 gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
     3661 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3721 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3781 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3841 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     3901 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     3961 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     4021 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     4081 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     4141 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4261 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4321 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4381 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4441 ttttggaggc ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca
     4501 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     4561 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4621 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4681 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4741 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4801 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4861 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4921 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4981 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5041 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5101 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5161 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5221 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5281 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5341 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5401 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     5461 gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc
     5521 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     6421 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     6481 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     6541 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     6601 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     6661 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     6721 gtccggcgta gaggatcgag atct
//
