LOCUS       sequence-471897-        5664 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express SLC35A2-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 5664)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SLC35A2-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5664
                     /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    2553..2674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2564..2583)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2789..2822)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2832..2879
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2911..3705
                     /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(2965..2984)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3575..3594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3922..4467
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4368..4387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4584..4601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4751..4799
                     /note="synthetic polyadenylation signal"
     CDS             complement(4823..5221)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5218..5227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5252..5609)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5266..5401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5418..5609
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5566..5586
                     /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 ggctgccgtg
     1201 ggcgcaggag gaagtacggc agcacctggt ccgggcgcgg tgtctgcagg cgctcttgaa
     1261 cctgggactg cttcagccgc ccaccgaaga ttgaagtata tttctcttgc tgttctcgta
     1321 gtgcaaaatg cttccctcat cctttctata agatacgctc ggacgttgcc gggcgatcga
     1381 tttttcgcaa caaccgcagt cgtcatggca gaggtcctta agggattgac gtgcctgctc
     1441 cttctctttg cgcaaaaacg aggaaatgta aaacacctgg ttctgtttct gcacgaggca
     1501 gtgcttgtcc aatatgttga cacactgaag ttggcggtac ccagcctcat ttatactttg
     1561 caaaataatc ttcaatatgt cgctatatca aatttgccgg cagctacgtt ccaagtcacc
     1621 taccaactga aaatcctgac aacggcattg ttctccgttc ttatgctgaa ccggtccttg
     1681 tcccgactcc aatgggccag tctccttctc ctctttacag gggtcgccat agttcaagct
     1741 caacaagcgg gaggaggagg tccccgccct ctcgatcaga atccgggagc cgggctcgcg
     1801 gctgtggtgg cctcctgcct tagctctggg ttcgcgggtg tatattttga gaagatcctg
     1861 aagggttctt ctggtagtgt ctggttgaga aatctgcagc ttggcctctt tgggacggcc
     1921 ctcgggttgg tgggtttgtg gtgggcggag ggcaccgcag tcgcaactcg cggattcttt
     1981 tttggataca caccagctgt ttggggtgtc gtattgaacc aagcatttgg cggattgctc
     2041 gtggcggttg ttgtgaaata cgcggacaac atactgaagg gattcgcgac ctccttgtcc
     2101 atcgtattgt ccaccgtcgc ctcaataaga ctcttcgggt ttcatgtaga tccgctcttt
     2161 gccttgggtg ctgggcttgt cataggagcg gtctatttgt actccctgcc tcggggagca
     2221 gcgaaagcca tagcctctgc ttccgcgtct gcctccggtc cgtgcgtcca tcagcaacct
     2281 ccgggacagc cgccgcctcc acagttgtcc tcccaccgcg gtgacctcat aaccgagccc
     2341 ttcctgccga aattgctgac caaagtgaag ggatccgttt ctcaaggcag ttcaggtggt
     2401 ggcgggagcg gaggtggagg ctcgagcggt gtgaccggct accggctgtt cgaggagatt
     2461 ctgtaaggcc gcgactctag agtcgacctg caggcatgca agctgatttg gctgctgcca
     2521 ccgctgagca aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     2581 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2641 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2701 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     2761 tggtaaaatc ttaattaaga attctaatat aacttcgtat agcatacatt atacgaagtt
     2821 atggatccac tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     2881 aggtgaggaa ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt
     2941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3061 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3421 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac
     3721 caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3781 caggggataa cgcaggaaag aacatgtcaa acatgagaat tacaacttat atcgtatggg
     3841 gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat
     3901 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     3961 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     4021 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     4081 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     4141 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     4201 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     4261 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     4321 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     4381 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     4441 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     4501 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     4561 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt
     4621 atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc
     4681 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt
     4741 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     4801 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     4861 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     4921 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     4981 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     5041 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     5101 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     5161 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     5221 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     5281 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     5341 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     5401 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5461 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5521 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5581 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5641 ttatttcagg ccatggtgct gcgc
//
