LOCUS       sequence-479825-        5038 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express CYBC1-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 5038)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5038)
  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            "Unpublished"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: CYBC1-LgBiT (TK) Vector
FEATURES             Location/Qualifiers
     source          1..5038
                     /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(1981..1999)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2036..2157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2047..2066)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2101..2120
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2402..2759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2402..2593
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2425..2445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2610..2745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2774..2821
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2853..3647
                     /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(2907..2926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3517..3536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3711..3759
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3998..4586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4078..4097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4702..4985
                     /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 agcggtgtct tcacactcga agatttcgtt
     1441 ggggactggg aacagacagc cgcctacaac ctggaccaag tccttgaaca gggaggtgtg
     1501 tccagtttgc tgcagaatct cgccgtgtcc gtaactccga tccaaaggat tgtccggagc
     1561 ggtgaaaatg ccctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcgcc
     1621 gaccaaatgg cccagatcga agaggtgttt aaggtggtgt accctgtgga tgatcatcac
     1681 tttaaggtga tcctgcccta tggcacactg gtaatcgacg gggttacgcc gaacatgctg
     1741 aactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta
     1801 acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcac ccccgacggc
     1861 tccatgctgt tccgagtaac catcaacagc taaggccgcg actctagagt cgacctgcag
     1921 gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1981 ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga
     2041 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2101 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2161 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     2221 agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac
     2281 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc
     2341 ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
     2401 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2761 agcttaatta actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     2821 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     2881 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2941 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3001 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3061 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3121 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3181 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3241 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3301 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3361 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3421 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3481 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3541 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3601 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3661 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat
     3721 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc
     3781 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     3841 agcaccgttt ctgcggactg gctttctacc cggtatcagc tcactcaaag gcggtaatac
     3901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4561 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     4621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4681 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     4741 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     4801 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     4861 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     4921 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     4981 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
