LOCUS       sequence-479823-        5746 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express CYBB-SmBiT 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 5746)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5746)
  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            "Unpublished"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: CYBB-SmBiT (TK) Vector
FEATURES             Location/Qualifiers
     source          1..5746
                     /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(2689..2707)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2744..2865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2755..2774)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2809..2828
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3110..3467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3110..3301
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3133..3153)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3318..3453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3482..3529
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3561..4355
                     /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(3615..3634)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4225..4244
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4419..4467
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4706..5294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4786..4805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5410..5693
                     /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 tagcgatcgc catggggaac tgggctgtga atgaggggct ctccattttt
      841 gtcattctgg tttggctggg gttgaacgtc ttcctctttg tctggtatta ccgggtttat
      901 gatattccac ctaagttctt ttacacaaga aaacttcttg ggtcagcact ggcactggcc
      961 agggcccctg cagcctgcct gaatttcaac tgcatgctga ttctcttgcc agtctgtcga
     1021 aatctgctgt ccttcctcag gggttccagt gcgtgctgct caacaagagt tcgaagacaa
     1081 ctggacagga atctcacctt tcataaaatg gtggcatgga tgattgcact tcactctgcg
     1141 attcacacca ttgcacatct atttaatgtg gaatggtgtg tgaatgcccg agtcaataat
     1201 tctgatcctt attcagtagc actctctgaa cttggagaca ggcaaaatga aagttatctc
     1261 aattttgctc gaaagagaat aaagaaccct gaaggaggcc tgtacctggc tgtgaccctg
     1321 ttggcaggca tcactggagt tgtcatcacg ctgtgcctca tattaattat cacttcctcc
     1381 accaaaacca tccggaggtc ttactttgaa gtcttttggt acacacatca tctctttgtg
     1441 atcttcttca ttggccttgc catccatgga gctgaacgaa ttgtacgtgg gcagaccgca
     1501 gagagtttgg ctgtgcataa tataacagtt tgtgaacaaa aaatctcaga atggggaaaa
     1561 ataaaggaat gcccaatccc tcagtttgct ggaaaccctc ctatgacttg gaaatggata
     1621 gtgggtccca tgtttctgta tctctgtgag aggttggtgc ggttttggcg atctcaacag
     1681 aaggtggtca tcaccaaggt ggtcactcac cctttcaaaa ccatcgagct acagatgaag
     1741 aagaaggggt tcaaaatgga agtgggacaa tacatttttg tcaagtgccc aaaggtgtcc
     1801 aagctggagt ggcacccttt tacactgaca tccgcccctg aggaagactt ctttagtatc
     1861 catatccgca tcgttgggga ctggacagag gggctgttca atgcttgtgg ctgtgataag
     1921 caggagtttc aagatgcgtg gaaactacct aagatagcgg ttgatgggcc ctttggcact
     1981 gccagtgaag atgtgttcag ctatgaggtg gtgatgttag tgggagcagg gattggggtc
     2041 acacccttcg catccattct caagtcagtc tggtacaaat attgcaataa cgccaccaat
     2101 ctgaagctca aaaagatcta cttctactgg ctgtgccggg acacacatgc ctttgagtgg
     2161 tttgcagatc tgctgcaact gctggagagc cagatgcagg aaaggaacaa tgccggcttc
     2221 ctcagctaca acatctacct cactggctgg gatgagtctc aggccaatca ctttgctgtg
     2281 caccatgatg aggagaaaga tgtgatcaca ggcctgaaac aaaagacttt gtatggacgg
     2341 cccaactggg ataatgaatt caagacaatt gcaagtcaac accctaatac cagaatagga
     2401 gttttcctct gtggacctga agccttggct gaaaccctga gtaaacaaag catctccaac
     2461 tctgagtctg gccctcgggg agtgcatttc attttcaaca aggaaaactt cgtttctcaa
     2521 ggcagttcag gtggtggcgg gagcggaggt ggaggctcga gcggtgtgac cggctaccgg
     2581 ctgttcgagg agattctgta aggccgcgac tctagagtcg acctgcaggc atgcaagctg
     2641 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     2701 aactagcata accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag
     2761 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2821 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2881 attcatttta tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac
     2941 ctctacaaat gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg
     3001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag
     3061 caccatggcc tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa
     3121 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaattaac
     3481 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     3541 ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3601 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3661 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3721 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3781 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3841 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     3901 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3961 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4021 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4081 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4141 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4201 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4261 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4321 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4381 cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat
     4441 tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg
     4501 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     4561 gcggactggc tttctacccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5281 aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5401 ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac
     5461 catgaaaaat ggcagcttca gtggattaag tgggggtaat gtggcctgta ccctctggtt
     5521 gcataggtat tcatacggtt aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt
     5581 ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt
     5641 acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac
     5701 cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
