LOCUS       sequence-479821-        6187 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express CYBB-LgBiT 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 6187)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6187)
  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            "Unpublished"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-LgBiT (TK) Vector
FEATURES             Location/Qualifiers
     source          1..6187
                     /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(3130..3148)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3185..3306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3196..3215)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3250..3269
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3551..3908
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3551..3742
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3574..3594)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3759..3894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3923..3970
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4002..4796
                     /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(4056..4075)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4666..4685
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4860..4908
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5147..5735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5227..5246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5851..6134
                     /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 gcggtgtctt cacactcgaa
     2581 gatttcgttg gggactggga acagacagcc gcctacaacc tggaccaagt ccttgaacag
     2641 ggaggtgtgt ccagtttgct gcagaatctc gccgtgtccg taactccgat ccaaaggatt
     2701 gtccggagcg gtgaaaatgc cctgaagatc gacatccatg tcatcatccc gtatgaaggt
     2761 ctgagcgccg accaaatggc ccagatcgaa gaggtgttta aggtggtgta ccctgtggat
     2821 gatcatcact ttaaggtgat cctgccctat ggcacactgg taatcgacgg ggttacgccg
     2881 aacatgctga actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     2941 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcacc
     3001 cccgacggct ccatgctgtt ccgagtaacc atcaacagct aaggccgcga ctctagagtc
     3061 gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     3121 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     3181 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3241 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3301 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     3361 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     3421 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3481 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     3541 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3601 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3661 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3781 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3841 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3901 tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3961 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4501 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4621 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4801 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca
     4861 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     4921 taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     4981 tccggggtca gcaccgtttc tgcggactgg ctttctaccc ggtatcagct cactcaaagg
     5041 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5101 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5161 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5221 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5281 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5341 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5401 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5461 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5521 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5581 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5641 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5701 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5821 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     5881 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     5941 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     6001 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     6061 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     6121 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     6181 gagatct
//
