LOCUS       sequence-471901-        4670 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express SmBiT-H4 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 4670)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4670)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SmBiT-H4 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..4670
                     /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    1733..1854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1744..1763)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1969..2002)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2012..2059
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2091..2885
                     /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(2145..2164)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2755..2774
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3043..3631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3123..3142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3757..3805
                     /note="synthetic polyadenylation signal"
     CDS             complement(3829..4227)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4224..4233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4258..4615)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4272..4407
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4424..4615
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4572..4592
                     /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 cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gtccggcaga ggaaagggcg gaaaaggctt aggcaaaggg
     1321 ggcgctaagc gccaccgcaa ggtcttgaga gacaacattc agggcatcac caagcctgcc
     1381 attcggcgtc tagctcggcg tggcggcgtt aagcggatct ctggcctcat ttacgaggag
     1441 acccgcggtg tgctgaaggt gttcctggag aatgtgattc gggacgcagt cacctacacc
     1501 gagcacgcca agcgcaagac cgtcacagcc atggatgtgg tgtacgcgct caagcgccag
     1561 gggcgcaccc tgtacggctt cggaggcgtt taaacgaatt cgggctcggt acccggggat
     1621 cctctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     1681 agctgagttg gctgctgcca ccgctgagca aggccgcttc gagcagacat gataagatac
     1741 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1801 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1861 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc
     1921 aagtaaaacc tctacaaatg tggtaaaatc ttaattaaga attctaatat aacttcgtat
     1981 agcatacatt atacgaagtt atggatccac tgttgacaat taatcatcgg catagtatat
     2041 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     2101 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2161 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2221 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2281 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2341 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2401 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2461 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2521 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2581 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2641 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2701 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2761 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2821 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2881 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     2941 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3481 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3601 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3721 gatcttcacc tagatccttt tatagtccgg aatcgtcaca caaaaaacca acacacagat
     3781 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagttt agccctccca
     3841 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     3901 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     3961 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     4021 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     4081 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     4141 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     4201 ttcttcttga gacaaaggct tggccatggt ggcgctagtg tcagaagaat cgagcttttt
     4261 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     4321 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     4381 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     4441 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     4501 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     4561 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcagaaggt
     4621 acctaaccaa gttcctcttt cagaggttat ttcaggccat ggtgctgcgc
//
