LOCUS       sequence-471898-        5723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SmBiT-SLC35A2 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 5723)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5723)
  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            "Unpublished"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-SLC35A2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5723
                     /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    2612..2733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2623..2642)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2848..2881)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2891..2938
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2970..3764
                     /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(3024..3043)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3634..3653
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3981..4526
                     /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     4427..4446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4643..4660
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4810..4858
                     /note="synthetic polyadenylation signal"
     CDS             complement(4882..5280)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5277..5286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5311..5668)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5325..5460
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5477..5668
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5625..5645
                     /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 ggctgccgtg ggcgcaggag gaagtacggc agcacctggt
     1321 ccgggcgcgg tgtctgcagg cgctcttgaa cctgggactg cttcagccgc ccaccgaaga
     1381 ttgaagtata tttctcttgc tgttctcgta gtgcaaaatg cttccctcat cctttctata
     1441 agatacgctc ggacgttgcc gggcgatcga tttttcgcaa caaccgcagt cgtcatggca
     1501 gaggtcctta agggattgac gtgcctgctc cttctctttg cgcaaaaacg aggaaatgta
     1561 aaacacctgg ttctgtttct gcacgaggca gtgcttgtcc aatatgttga cacactgaag
     1621 ttggcggtac ccagcctcat ttatactttg caaaataatc ttcaatatgt cgctatatca
     1681 aatttgccgg cagctacgtt ccaagtcacc taccaactga aaatcctgac aacggcattg
     1741 ttctccgttc ttatgctgaa ccggtccttg tcccgactcc aatgggccag tctccttctc
     1801 ctctttacag gggtcgccat agttcaagct caacaagcgg gaggaggagg tccccgccct
     1861 ctcgatcaga atccgggagc cgggctcgcg gctgtggtgg cctcctgcct tagctctggg
     1921 ttcgcgggtg tatattttga gaagatcctg aagggttctt ctggtagtgt ctggttgaga
     1981 aatctgcagc ttggcctctt tgggacggcc ctcgggttgg tgggtttgtg gtgggcggag
     2041 ggcaccgcag tcgcaactcg cggattcttt tttggataca caccagctgt ttggggtgtc
     2101 gtattgaacc aagcatttgg cggattgctc gtggcggttg ttgtgaaata cgcggacaac
     2161 atactgaagg gattcgcgac ctccttgtcc atcgtattgt ccaccgtcgc ctcaataaga
     2221 ctcttcgggt ttcatgtaga tccgctcttt gccttgggtg ctgggcttgt cataggagcg
     2281 gtctatttgt actccctgcc tcggggagca gcgaaagcca tagcctctgc ttccgcgtct
     2341 gcctccggtc cgtgcgtcca tcagcaacct ccgggacagc cgccgcctcc acagttgtcc
     2401 tcccaccgcg gtgacctcat aaccgagccc ttcctgccga aattgctgac caaagtgaag
     2461 ggatccgttt aaacgaattc gggctcggta cccggggatc ctctagagtc gacctgcagg
     2521 catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     2581 cgctgagcaa ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2641 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2701 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2761 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     2821 ggtaaaatct taattaagaa ttctaatata acttcgtata gcatacatta tacgaagtta
     2881 tggatccact gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     2941 ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt
     3001 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3061 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3121 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3181 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3241 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3301 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3361 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3421 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3481 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3541 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3601 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3661 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3721 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgattcgaa atgaccgacc
     3781 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3841 aggggataac gcaggaaaga acatgtcaaa catgagaatt acaacttata tcgtatgggg
     3901 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     3961 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     4021 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     4081 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     4141 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     4201 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4261 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     4321 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     4381 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     4441 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     4501 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     4561 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     4621 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     4681 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     4741 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     4801 cggaatcgtc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
     4861 ggccatcgtg atggctagag tttagccctc ccacacataa ccagagggca gcaattcacg
     4921 aatcccaact gccgtcggct gtccatcact gtccttcact atggctttga tcccaggatg
     4981 cagatcgaga agcacctgtc ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt
     5041 tccgatcgcg acgatacaag tcaggttgcc agctgccgca gcagcagcag tgcccagcac
     5101 cacgagttct gcacaaggtc ccccagtaaa atgatataca ttgacaccag tgaagatgcg
     5161 gccgtcgcta gagagagctg cgctggcgac gctgtagtct tcagagatgg ggatgctgtt
     5221 gattgtagcc gttgctcttt caatgagggt ggattcttct tgagacaaag gcttggccat
     5281 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     5341 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     5401 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5461 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5521 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     5581 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     5641 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     5701 tatttcaggc catggtgctg cgc
//
