LOCUS       sequence-473316-        5471 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express LgBiT-TUBEs 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 5471)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5471)
  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: LgBiT-TUBEs Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5471
                     /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"
     primer_bind     275..293
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(1525..1543)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1526..1543)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1663..1966
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2121..2141
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2174..2221
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2253..3047
                     /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(2307..2326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2917..2936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3205..3793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3285..3304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3919..3967
                     /note="synthetic polyadenylation signal"
     CDS             complement(3991..5028)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4004..4012
                     /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(5059..5416)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5073..5208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5225..5416
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5373..5393
                     /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 ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttattagcct gtcagcaaat
      421 actcgacagc tcgatgagga ttgttatagg aggcccgcag ggcagctaca acgcgctcgc
      481 gttcgtaacc cattgacatt atctcagtaa gcatagtctc gtattcggat ccgctgccac
      541 ttccgctccc accggtcagc aggtattcaa ccgccctatg cggattatta tatgaggctc
      601 gcagcgcagc gaccactctc tccctctcgt atcccatact cataatctcg gtgagcatcg
      661 tctcatattc actgccgcta cctgatccac taccacccgt cagcagatac tcaacagctc
      721 tatgggggtt attataggaa gctcggagcg ctgcgacaac tcgttcccgt tcatagccca
      781 tggacataat ctctgtcagc atggtctcat actctgaacc tgacccgctt ccactccctc
      841 ctgtcagaag atattccacc gctcggtgag gattgttgta gctcgcgcgc agagccgcca
      901 caacgcgctc ccgttcatat cccattgaca ttatttccgt gagcatggtt tcatattcgg
      961 agccagaacc tcctcccccg gatcctcctc cgccggaccc tccaccacca ctgttgatgg
     1021 ttactcggaa cagcatggag ccgtcggggg tgatcaggcg ctcgtcgata attttgttgc
     1081 cgttccacag ggtccctgtt acagtgatct ttttgccgtc gaacacggcg atgccttcat
     1141 acggccgtcc gaaatagttc agcatgttcg gcgtaacccc gtcgattacc agtgtgccat
     1201 agggcaggat caccttaaag tgatgatcat ccacagggta caccacctta aacacctctt
     1261 cgatctgggc catttggtcg gcgctcagac cttcatacgg gatgatgaca tggatgtcga
     1321 tcttcagggc attttcaccg ctccggacaa tcctttggat cggagttacg gacacggcga
     1381 gattctgcag caaactggac acacctccct gttcaaggac ttggtccagg ttgtaggcgg
     1441 ctgtctgttc ccagtcccca acgaaatctt cgagtgtgaa gaccatggtg agcgctagct
     1501 gcaaaaagaa caagtagctt gtattctata gtgtcaccta aatgggcgat ctgacggttc
     1561 actaaaccag ctctgcttat ataggcctcc ctggcgcgcc actagttatt aatagtaatc
     1621 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1681 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1741 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1801 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     1861 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     1921 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     1981 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2041 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2101 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtcaaatc
     2161 gaattctaat actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     2221 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     2281 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2341 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2401 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2461 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2521 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2581 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2641 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2701 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2761 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2821 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2881 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2941 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3001 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     3061 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3121 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3181 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3241 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3301 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3361 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3421 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3481 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3541 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3601 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     3661 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3721 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3781 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3841 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3901 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     3961 ttttattgcg gccatcgtga tggctagagt ctaaactcga cctacctcct tagcgcgcgg
     4021 tcgtgtactg ggccggcggt tgccgctgtc ggccagcacc tcgacgcagc cgtcggtcca
     4081 tacggctgcg ctccggcgag cgatttgtgt gcgaccgacg gtgccggccc cgctgcggac
     4141 gatggcatcg cagcggcctt gagcccaggc agcatcgtcg aagttgccgt ccacgaggct
     4201 ctggtagagc tgatccaggc cgatgcgcag catgtaggct cgcagacgag ggctgccggc
     4261 cagctcggga tgccggcgct cgaagtagcg agtctgctgc tccatgcaag ccagccaggg
     4321 ccgccagaag aagatgttgg ccacctcgta ctgactgtcc ccgaacatag cttcggacca
     4381 gtcgattacg gcggtgatgc ggccgttgtc ggtcaggacg ttgttgctgc cgaagtcggc
     4441 atggacgagg tggcgcacct cgggacagtc ttcggcccac agcatcagtt cgtccagggc
     4501 ttgagctacg ctggcggaca cggtgtcgtc catcacggtc tgccagtggt agacatgggg
     4561 atcagcaatg gcgcaaatga aatcccgcca ggttgtgtac tggccgatgc cttggggccc
     4621 gaatgggccg aagccgctgg tttgactgag gtcggcggcg gcaatagcat ccatagcttc
     4681 ggcgacaggc tgtaacacag caggcagctc tgtttcgggg aggtcttgga gagtaacgcc
     4741 ttgggcgcgt ctactgatgc agtatgtcag gctctcgctg aactcgccga tgtccaacac
     4801 ttcggggatg ggtagtgcag cgctggcgaa gtggcggtac acgtagcggt ctttgtagaa
     4861 gccatcagcg cagctattca cccgcagtac atagccgcgt ccgccgacat cgaagctgaa
     4921 ggctcggctc tcttcgccct ccgacaactg catcaggtcg ctcacactgt cgaacttctc
     4981 gatgagaaat ttttcaacgc tggtagcggt gagttcgggc ttcttcatgg tggcgctagt
     5041 gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     5101 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     5161 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     5221 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5281 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     5341 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
     5401 gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta tttcaggcca
     5461 tggtgctgcg c
//
