LOCUS       sequence-474107-        5459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SmBiT-NCK1 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 5459)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5459)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: SmBiT-NCK1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(747..1541)
                     /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(858..877)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1468..1487
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1573..1620)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    1630..1663
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3245..3264)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3246..3264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3306..3325)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3308..3440
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(3576..3779)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3605..3625)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3780..4083
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4316..4397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4476..4833
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4476..4667
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4499..4519)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4684..4819
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4858..4867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4864..5262
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5286..5334
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaatcag aagaactcgt caagaaggcg atagaaggcg
      781 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
      841 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      901 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      961 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     1021 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1081 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1141 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1201 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1261 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1321 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1381 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1441 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1501 gcggccggag aacctgcgtg caatccatct tgttcaatca tgatctgcct cctgggttta
     1561 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     1621 agtggatcca taacttcgta taatgtatgc tatacgaagt tatattagaa ttcttaatta
     1681 agattttacc acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct
     1741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc
     1921 ttgctcagcg gtggcagcag ccaaatcagc ttgcatgcct gcaggtcgac tctagaggat
     1981 ccccgggtac cgagcccgaa ttcgtttaaa ctgataaatg cttgacaaga tataattttt
     2041 ctccttgttc acttgtaaaa attggtgcct ttttgtaatg ttctacaagt tcttccatgg
     2101 tgctgaattt acgctgccca atgcagtaga cagtctcttt tagttggact ttaaaatgct
     2161 tgtttttccc ttgtgctttt agtgatactg agaaatcatt tggcgaagat tcactatcac
     2221 gaatgaggaa atccccttca tgtcctcttt catttaatgc catttctgct tgatgcctgg
     2281 tgactttgcc ataataccaa ggattgccag caaactttcc agtgagtgaa ggcctaatgt
     2341 aatcacactg tggaggtgat ggttccaaac ctgaagttaa tggattattc tgcataacgg
     2401 taacatagtt ttttggtact agaccaacca taccattgat cttcctgcat ttccaccact
     2461 ctgggtcatt ttcaggtttt tcaataacat ccattacatc tcctttctcg aaattaagtt
     2521 cttcatcatt agatgagctg aatgggtaaa gagcctgtac cacatgcaac acttgcccag
     2581 tatttaggtt attgacgact gctgctaatt tctctgacag agaacccaca tggtcaccca
     2641 aaggactgtc accttcttca gttacatagt ttgaagggaa ccatccaact tgtccattgt
     2701 agctaccacg ccaccaccca tcactgcatt tctccatgac gatcaccttt gtccccttta
     2761 tcaatgataa ttcatcctct ctctcagcca tgtagttaaa tttcacataa gcgggcatgt
     2821 tgaggtcata gagacgttcc cctgggtcaa caaaactatc atcagcagga gatgcagaat
     2881 ctggcacact aggttttctt ttcacttttc caatgcctaa ggtatccttt aggtttttca
     2941 caatagatgc tttccgagca ctgtttttcc tttccacata gttagaaggc acaaaacctg
     3001 ttttattcat ggaatttcga actcgccacc aggacttaga atcatccaga agccataatc
     3061 tctcattctt cttgatgtcc aactcttgtt cttgttgggc cacataatca aatttggcta
     3121 ctaccaccac ttcttctgcc atggcgatcg caccgctcga gcctccacct ccgctcccgc
     3181 caccaccgga gctcccgaga atctcctcga acagccggta gccggtcacc atggtgagcg
     3241 ctagccctat agtgagtcgt attaatactc tagccttaag agctgtaatt gaactgggag
     3301 tggacacctg tggagagaaa ggcaaagtgg atgtcagtaa gaccaatagg tgcctatcag
     3361 aaacgcaaga gtcttctctg tctcgacaag cccagtttct attggtctcc ttaaacctgt
     3421 cttgtaacct tgatacttac ctgcccagtg cctcacgacc aacttctgca gcttaagttc
     3481 gagactgttg tgtcagaagc actgactgcg ttagcaattt aactgtgata aactaccgca
     3541 ataaagcttc tagtgatctg acggttcact aaaccagctc tgcttatata gacctcccac
     3601 cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca ttttggaaag
     3661 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     3721 aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc
     3781 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc
     3841 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga
     3901 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     3961 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     4021 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     4081 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     4141 ctagtggcgc gccataatcc taggctctcg tacgaagtca gctgataacg tcgacgtatt
     4201 gcggccgcgg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4261 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     4321 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4381 agcatttttt tcactgccga cgtcttaatt aacgcgtctt gcgcagcacc atggcctgaa
     4441 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     4501 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4561 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4621 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4681 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4741 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4801 ggaggctttt ttggaggcct aggcttttgc aaaaagctcg attcttctga cactagcgcc
     4861 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     4921 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     4981 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     5041 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     5101 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     5161 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     5221 attcgtgaat tgctgccctc tggttatgtg tgggagggct aaactctagc catcacgatg
     5281 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgacgatt
     5341 ccggactata aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5401 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
