LOCUS       sequence-474677-        6472 bp    DNA     circular SYN 05-JUN-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 6472)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6472)
  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: NanoBiT Assay Vector, HER1(1-668)-LgBiT
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..286
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     243..263
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    365..446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(649..1098)
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     promoter        complement(1668..1686)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1669..1686)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1806..2109
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2110..2313
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2264..2284
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          2449..2581
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2564..2583
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     2625..2644
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2625..2643
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3876..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3887..3906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4112..4145)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4155..4202
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4234..5028
                     /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(4288..4307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4898..4917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5186..5774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5266..5285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5900..5948
                     /note="synthetic polyadenylation signal"
     CDS             complement(5972..6370)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6367..6376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(6401..6472,1..286))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6415..6472,1..78)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
       61 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
      121 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
      181 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      241 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
      301 agttcctctt tcagaggtta tttcaggcca tggtgctgcg caagacgcgt taattaagac
      361 gtcggcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      421 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      481 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
      541 ctgattatga tccgcggccg caatacgtcg acgttatcag ctgacttcgt acgagagcct
      601 aggtgtttct agaggatccc cgggtaccga gcccgaattc gtttaaactc attttgcagt
      661 catcatctgt acgaattctt catagttgac ttgtccgtct ccatcaatat ctgcttctct
      721 gatcatttca tctacttctt catctgttag tttttctcct aagtttgtca tgacgtgacg
      781 tagttctgct gcactgatat aaccattgcc atccttgtca aagactcgga atgcctcacg
      841 gatttcttct tcactatctg tatctttcat ttttctagcc atcatagtca aaaattcggg
      901 gaagtcaatg gtgccattac catcagcatc cacttcattg atcatatcct gcaattcagc
      961 ttctgttggg ttctgaccca gtgacctcat gacagttcca agttcctttg ttgtgatggt
     1021 gccatcgcca tctttatcaa atagggagaa ggcttccttg aattcagcaa tctgttcttc
     1081 ggtcagctga tcagccatgg cgatcgcacc gctcgagcct ccacctccgc tcccgccacc
     1141 accggagctc ccactgttga tggttactcg gaacagcatg gagccgtcgg gggtgatcag
     1201 gcgctcgtcg ataattttgt tgccgttcca cagggtccct gttacagtga tctttttgcc
     1261 gtcgaacacg gcgatgcctt catacggccg tccgaaatag ttcagcatgt tcggcgtaac
     1321 cccgtcgatt accagtgtgc catagggcag gatcacctta aagtgatgat catccacagg
     1381 gtacaccacc ttaaacacct cttcgatctg ggccatttgg tcggcgctca gaccttcata
     1441 cgggatgatg acatggatgt cgatcttcag ggcattttca ccgctccgga caatcctttg
     1501 gatcggagtt acggacacgg cgagattctg cagcaaactg gacacacctc cctgttcaag
     1561 gacttggtcc aggttgtagg cggctgtctg ttcccagtcc ccaacgaaat cttcgagtgt
     1621 gaagaccatg gtgagcgcta gctgcaaaaa gaacaagtag cttgtattct atagtgtcac
     1681 ctaaatgggc gatctgacgg ttcactaaac cagctctgct tatataggcc tccctggcgc
     1741 gccactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     1801 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     1861 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     1921 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     1981 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     2041 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     2101 attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca
     2161 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     2221 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
     2281 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcactaga
     2341 agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca
     2401 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
     2461 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     2521 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     2581 ggtgtccact cccagttcaa ttacagctct taaggctaga gtattaatac gactcactat
     2641 agggctagcg ctcaccatgg tgaccggcta ccggctgttc gaggagattc tcgggagctc
     2701 cggtggtggc gggagcggag gtggaggctc gagcggtgcg atcgccatgt atggggtctt
     2761 tgtgcggatt ctgactgagc gttcacagag gaaggcgttc ctgcaggccc ggagctgcat
     2821 tgaggaccga ctgaggctgg aggatgagaa cgagaagcag gagcggctcc tcatgagcct
     2881 cctgccccgg aacgttgcca tggagatgaa ggaggacttc ctgaagcccc ctgagaggat
     2941 tttccacaag atttacatcc agaggcacga caatgtgagc atcctgtttg ctgacatcgt
     3001 gggtttcacg ggcttggcat cccagtgcac agcccaggag ctggtgaaac tcctcaatga
     3061 gctcttcggc aagttcgatg aattagccac ggagaaccac tgtcgccgca tcaagattct
     3121 cggggactgc tactactgcg tgtcgggcct cacccagccc aagactgacc atgcccactg
     3181 ctgtgtggag atgggactcg acatgattga taccatcaca tctgtggctg aagccaccga
     3241 ggtggatctg aacatgcgtg tgggtctgca cacgggcagg gtcctctgtg gtgtcctggg
     3301 cttgcgcaag tggcagtacg acgtgtggtc caatgatgtg accttggcca atgtcatgga
     3361 agccgctggc ctgccaggga aggttcatat cacaaagacg accctagcgt gcttgaatgg
     3421 ggactacgag gtagaaccgg gttacggaca tgagaggaac agtttcttga aaactcataa
     3481 catcgaaacc ttttttattg tgccatccca tcgccgaaag atatttccag gcctgattct
     3541 ctcagatata aaaccggcca aaaggatgaa gttcaagact gtctgctacc tgctggtgca
     3601 gctcatgcac tgccggaaaa tgttcaaggc cgagatcccc ttctccaatg tcatgacctg
     3661 cgaggacgat gacaaggttc agatccatcc ctcactgagt ggcctggatg tcctcctggg
     3721 gttttggtct cccaccctgc aacggggacg actgtgcacg tgagtttaaa cgaattcggg
     3781 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat ttggctgctg
     3841 ccaccgctga gcaaggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4021 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     4081 atgtggtaaa atcttaatta agaattctaa tataacttcg tatagcatac attatacgaa
     4141 gttatggatc cactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     4201 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     4261 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4321 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4381 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4441 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4501 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4561 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4621 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4681 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4741 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4801 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4861 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4921 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4981 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaaatgacc
     5041 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5101 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5161 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5221 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5281 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5341 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5401 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5461 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5521 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5581 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5641 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5701 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5761 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5821 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5881 ttttatagtc cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat
     5941 attttattgc ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca
     6001 gcaattcacg aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga
     6061 tcccaggatg cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc
     6121 tgttctcatt tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag
     6181 tgcccagcac cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag
     6241 tgaagatgcg gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg
     6301 ggatgctgtt gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag
     6361 gcttggccat ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc
     6421 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tc
//
