LOCUS       sequence-486459-        4597 bp    DNA     circular SYN 21-SEP-2025
DEFINITION  Express CYBC1-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 4597)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4597)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: CYBC1-SmBiT (TK) Vector
FEATURES             Location/Qualifiers
     source          1..4597
                     /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(1540..1558)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1595..1716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1606..1625)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1660..1679
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1961..2318
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1961..2152
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1984..2004)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2169..2304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2333..2380
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2412..3206
                     /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(2466..2485)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3076..3095
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3270..3318
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3557..4145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3637..3656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4261..4544
                     /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 catgtacctg caggtggaga cccgcaccag ctcccgcctc
      841 catctgaaga gggctccagg catccggtcc tggtccctgc tggttggaat cttgtcgatt
      901 ggcctggctg ctgcctacta cagcggagat agcctgggct ggaagctctt ctacgtcaca
      961 ggctgcctgt ttgtggctgt gcagaacttg gaggactggg aggaagccat cttcgacaag
     1021 agcacaggga aggttgtttt gaagacgttc agcctctaca agaagctgct gactcttttc
     1081 agagctggcc acgaccaggt ggtggtcctg ctccatgatg tccgtgatgt gagcgtggag
     1141 gaggagaagg tccggtactt cgggaaaggc tacatggtgg tgctccggct tgcgacgggc
     1201 ttctcccacc ccctcacgca gagtgcagtc atgggccacc gcagtgatgt ggaagccatc
     1261 gccaagctca tcaccagctt cctggagctg cactgccttg agagccccac agagctgtct
     1321 cagagcagcg acagtgaggc cggtgaccct gcaagccaga gcgtttctca aggcagttca
     1381 ggtggtggcg ggagcggagg tggaggctcg agcggtgtga ccggctaccg gctgttcgag
     1441 gagattctgt aaggccgcga ctctagagtc gacctgcagg catgcaagct gatccggctg
     1501 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1561 aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     1621 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1681 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1741 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     1801 tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     1861 ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc
     1921 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     1981 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2041 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2161 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2221 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2281 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa
     2341 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     2401 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2461 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2521 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2581 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2641 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2701 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     2761 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     2821 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     2881 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     2941 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3001 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3061 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3121 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3181 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3241 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     3301 gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc
     3361 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     3421 ctttctaccc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3481 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3541 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3601 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3661 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3721 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3781 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3841 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3901 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3961 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4021 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4081 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4141 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4201 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     4261 ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa
     4321 tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta
     4381 ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca
     4441 tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag
     4501 ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact
     4561 ccggccacga tgcgtccggc gtagaggatc gagatct
//
