LOCUS       sequence-471895-        6152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SLC35A2-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 6152)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6152)
  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            "Unpublished"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: SLC35A2-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2032..2050)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2033..2050)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2170..2473
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2628..2648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2681..2728
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2760..3554
                     /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(2814..2833)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3424..3443
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3771..4316
                     /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     4217..4236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4433..4450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4600..4648
                     /note="synthetic polyadenylation signal"
     CDS             complement(4672..5709)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4685..4693
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(5740..6097)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5754..5889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5906..6097
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6054..6074
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaacggatc ccttcacttt ggtcagcaat ttcggcagga
      841 agggctcggt tatgaggtca ccgcggtggg aggacaactg tggaggcggc ggctgtcccg
      901 gaggttgctg atggacgcac ggaccggagg cagacgcgga agcagaggct atggctttcg
      961 ctgctccccg aggcagggag tacaaataga ccgctcctat gacaagccca gcacccaagg
     1021 caaagagcgg atctacatga aacccgaaga gtcttattga ggcgacggtg gacaatacga
     1081 tggacaagga ggtcgcgaat cccttcagta tgttgtccgc gtatttcaca acaaccgcca
     1141 cgagcaatcc gccaaatgct tggttcaata cgacacccca aacagctggt gtgtatccaa
     1201 aaaagaatcc gcgagttgcg actgcggtgc cctccgccca ccacaaaccc accaacccga
     1261 gggccgtccc aaagaggcca agctgcagat ttctcaacca gacactacca gaagaaccct
     1321 tcaggatctt ctcaaaatat acacccgcga acccagagct aaggcaggag gccaccacag
     1381 ccgcgagccc ggctcccgga ttctgatcga gagggcgggg acctcctcct cccgcttgtt
     1441 gagcttgaac tatggcgacc cctgtaaaga ggagaaggag actggcccat tggagtcggg
     1501 acaaggaccg gttcagcata agaacggaga acaatgccgt tgtcaggatt ttcagttggt
     1561 aggtgacttg gaacgtagct gccggcaaat ttgatatagc gacatattga agattatttt
     1621 gcaaagtata aatgaggctg ggtaccgcca acttcagtgt gtcaacatat tggacaagca
     1681 ctgcctcgtg cagaaacaga accaggtgtt ttacatttcc tcgtttttgc gcaaagagaa
     1741 ggagcaggca cgtcaatccc ttaaggacct ctgccatgac gactgcggtt gttgcgaaaa
     1801 atcgatcgcc cggcaacgtc cgagcgtatc ttatagaaag gatgagggaa gcattttgca
     1861 ctacgagaac agcaagagaa atatacttca atcttcggtg ggcggctgaa gcagtcccag
     1921 gttcaagagc gcctgcagac accgcgcccg gaccaggtgc tgccgtactt cctcctgcgc
     1981 ccacggcagc catggcgatc gctagctgca aaaagaacaa gtagcttgta ttctatagtg
     2041 tcacctaaat gggcgatctg acggttcact aaaccagctc tgcttatata ggcctccctg
     2101 gcgcgccact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2161 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2221 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2281 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2341 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2401 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     2461 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     2521 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2581 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     2641 taggcgtgta cggtgggagg tcaaatcgaa ttctaatact gttgacaatt aatcatcggc
     2701 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     2761 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2821 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2881 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2941 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3001 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3061 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3121 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3181 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3241 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3301 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3361 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3421 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3481 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3541 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     3601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtcaaa
     3661 catgagaatt acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata
     3721 aattgcactg aaatctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt
     3781 tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt
     3841 tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag
     3901 tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc
     3961 tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga
     4021 ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat
     4081 acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca
     4141 aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca
     4201 cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4261 actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg
     4321 gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc
     4381 tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc
     4441 agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc
     4501 cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa
     4561 gccagtatac actccgctag cgctgatgtc cggaatcgtc acacaaaaaa ccaacacaca
     4621 gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag tctaaactcg
     4681 acctacctcc ttagcgcgcg gtcgtgtact gggccggcgg ttgccgctgt cggccagcac
     4741 ctcgacgcag ccgtcggtcc atacggctgc gctccggcga gcgatttgtg tgcgaccgac
     4801 ggtgccggcc ccgctgcgga cgatggcatc gcagcggcct tgagcccagg cagcatcgtc
     4861 gaagttgccg tccacgaggc tctggtagag ctgatccagg ccgatgcgca gcatgtaggc
     4921 tcgcagacga gggctgccgg ccagctcggg atgccggcgc tcgaagtagc gagtctgctg
     4981 ctccatgcaa gccagccagg gccgccagaa gaagatgttg gccacctcgt actgactgtc
     5041 cccgaacata gcttcggacc agtcgattac ggcggtgatg cggccgttgt cggtcaggac
     5101 gttgttgctg ccgaagtcgg catggacgag gtggcgcacc tcgggacagt cttcggccca
     5161 cagcatcagt tcgtccaggg cttgagctac gctggcggac acggtgtcgt ccatcacggt
     5221 ctgccagtgg tagacatggg gatcagcaat ggcgcaaatg aaatcccgcc aggttgtgta
     5281 ctggccgatg ccttggggcc cgaatgggcc gaagccgctg gtttgactga ggtcggcggc
     5341 ggcaatagca tccatagctt cggcgacagg ctgtaacaca gcaggcagct ctgtttcggg
     5401 gaggtcttgg agagtaacgc cttgggcgcg tctactgatg cagtatgtca ggctctcgct
     5461 gaactcgccg atgtccaaca cttcggggat gggtagtgca gcgctggcga agtggcggta
     5521 cacgtagcgg tctttgtaga agccatcagc gcagctattc acccgcagta catagccgcg
     5581 tccgccgaca tcgaagctga aggctcggct ctcttcgccc tccgacaact gcatcaggtc
     5641 gctcacactg tcgaacttct cgatgagaaa tttttcaacg ctggtagcgg tgagttcggg
     5701 cttcttcatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     5761 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5821 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5881 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     5941 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     6001 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     6061 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     6121 ttcagaggtt atttcaggcc atggtgctgc gc
//
