LOCUS       sequence-479822-        6213 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express LgBiT-CYBB Fusion Protein in Mammalian Cells under a TK 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6213)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LgBiT-CYBB (TK) Vector
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..753
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(3156..3174)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3211..3332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3222..3241)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3276..3295
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3577..3934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3577..3768
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3600..3620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3785..3920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3949..3996
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4028..4822
                     /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(4082..4101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4692..4711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4886..4934
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5173..5761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5253..5272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5877..6160
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 aaatgagtct tcggacctcg cgggggccgc ttaagcggtg gttagggttt gtctgacgcg
       61 gggggagggg gaaggaacga aacactctca ttcggaggcg gctcggggtt tggtcttggt
      121 ggccacgggc acgcagaaga gcgccgcgat cctcttaagc acccccccgc cctccgtgga
      181 ggcgggggtt tggtcggcgg gtggtaactg gcgggccgct gactcgggcg ggtcgcgcgc
      241 cccagagtgt gaccttttcg gtctgctcgc agacccccgg gcggcgccgc cgcggcggcg
      301 acgggctcgc tgggtcctag gctccatggg gaccgtatac gtggacaggc tctggagcat
      361 ccgcacgact gcggtgatat taccggagac cttctgcggg acgagccggg tcacgcggct
      421 gacgcggagc gtccgttggg cgacaaacac caggacgggg cacaggtaca ctatcttgtc
      481 acccggaggc gcgagggact gcaggagctt cagggagtgg cgcagctgct tcatccccgt
      541 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
      601 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaagtcga acacgcagat
      661 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
      721 gaacaccgag cgaccctgca gcgacccgct taaaagcttg gcaatccggt actgttggta
      781 aagccaccgc tagcgctcac catggtcttc acactcgaag atttcgttgg ggactgggaa
      841 cagacagccg cctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgctg
      901 cagaatctcg ccgtgtccgt aactccgatc caaaggattg tccggagcgg tgaaaatgcc
      961 ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcgccga ccaaatggcc
     1021 cagatcgaag aggtgtttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc
     1081 ctgccctatg gcacactggt aatcgacggg gttacgccga acatgctgaa ctatttcgga
     1141 cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg
     1201 tggaacggca acaaaattat cgacgagcgc ctgatcaccc ccgacggctc catgctgttc
     1261 cgagtaacca tcaacagtgg gagctccggt ggtggcggga gcggaggtgg aggctcgagc
     1321 ggtgcgatcg ccatggggaa ctgggctgtg aatgaggggc tctccatttt tgtcattctg
     1381 gtttggctgg ggttgaacgt cttcctcttt gtctggtatt accgggttta tgatattcca
     1441 cctaagttct tttacacaag aaaacttctt gggtcagcac tggcactggc cagggcccct
     1501 gcagcctgcc tgaatttcaa ctgcatgctg attctcttgc cagtctgtcg aaatctgctg
     1561 tccttcctca ggggttccag tgcgtgctgc tcaacaagag ttcgaagaca actggacagg
     1621 aatctcacct ttcataaaat ggtggcatgg atgattgcac ttcactctgc gattcacacc
     1681 attgcacatc tatttaatgt ggaatggtgt gtgaatgccc gagtcaataa ttctgatcct
     1741 tattcagtag cactctctga acttggagac aggcaaaatg aaagttatct caattttgct
     1801 cgaaagagaa taaagaaccc tgaaggaggc ctgtacctgg ctgtgaccct gttggcaggc
     1861 atcactggag ttgtcatcac gctgtgcctc atattaatta tcacttcctc caccaaaacc
     1921 atccggaggt cttactttga agtcttttgg tacacacatc atctctttgt gatcttcttc
     1981 attggccttg ccatccatgg agctgaacga attgtacgtg ggcagaccgc agagagtttg
     2041 gctgtgcata atataacagt ttgtgaacaa aaaatctcag aatggggaaa aataaaggaa
     2101 tgcccaatcc ctcagtttgc tggaaaccct cctatgactt ggaaatggat agtgggtccc
     2161 atgtttctgt atctctgtga gaggttggtg cggttttggc gatctcaaca gaaggtggtc
     2221 atcaccaagg tggtcactca ccctttcaaa accatcgagc tacagatgaa gaagaagggg
     2281 ttcaaaatgg aagtgggaca atacattttt gtcaagtgcc caaaggtgtc caagctggag
     2341 tggcaccctt ttacactgac atccgcccct gaggaagact tctttagtat ccatatccgc
     2401 atcgttgggg actggacaga ggggctgttc aatgcttgtg gctgtgataa gcaggagttt
     2461 caagatgcgt ggaaactacc taagatagcg gttgatgggc cctttggcac tgccagtgaa
     2521 gatgtgttca gctatgaggt ggtgatgtta gtgggagcag ggattggggt cacacccttc
     2581 gcatccattc tcaagtcagt ctggtacaaa tattgcaata acgccaccaa tctgaagctc
     2641 aaaaagatct acttctactg gctgtgccgg gacacacatg cctttgagtg gtttgcagat
     2701 ctgctgcaac tgctggagag ccagatgcag gaaaggaaca atgccggctt cctcagctac
     2761 aacatctacc tcactggctg ggatgagtct caggccaatc actttgctgt gcaccatgat
     2821 gaggagaaag atgtgatcac aggcctgaaa caaaagactt tgtatggacg gcccaactgg
     2881 gataatgaat tcaagacaat tgcaagtcaa caccctaata ccagaatagg agttttcctc
     2941 tgtggacctg aagccttggc tgaaaccctg agtaaacaaa gcatctccaa ctctgagtct
     3001 ggccctcggg gagtgcattt cattttcaac aaggaaaact tcgtttaaac gaattcgggc
     3061 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     3121 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3181 ccttggggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3241 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3301 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3361 ttcaggttca gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg
     3421 gtaaaatcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     3481 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     3541 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     3601 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3721 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3781 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3841 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3901 aggaggcttt tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa
     3961 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag
     4021 gcagatcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4201 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4501 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4681 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4801 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4861 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4921 tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt
     4981 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     5041 ctacccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5101 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5161 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5221 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5281 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5341 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5401 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5461 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5521 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5581 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5641 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5701 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     5761 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5821 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     5881 aaatacagga acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc
     5941 agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca
     6001 tacggttaaa atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt
     6061 tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat
     6121 tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg
     6181 ccacgatgcg tccggcgtag aggatcgaga tct
//
