LOCUS       sequence-479827-        4623 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express SmBiT-CYBC1 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 4623)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4623)
  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            "Unpublished"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: SmBiT-CYBC1 (TK) Vector
FEATURES             Location/Qualifiers
     source          1..4623
                     /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(1566..1584)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1621..1742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1632..1651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1686..1705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1987..2344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1987..2178
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2010..2030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2195..2330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2359..2406
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2438..3232
                     /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(2492..2511)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3102..3121
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3296..3344
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3583..4171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3663..3682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4287..4570
                     /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 catggtgacc ggctaccggc tgttcgagga gattctcggg
      841 agctccggtg gtggcgggag cggaggtgga ggctcgagcg gtgcgatcgc catgtacctg
      901 caggtggaga cccgcaccag ctcccgcctc catctgaaga gggctccagg catccggtcc
      961 tggtccctgc tggttggaat cttgtcgatt ggcctggctg ctgcctacta cagcggagat
     1021 agcctgggct ggaagctctt ctacgtcaca ggctgcctgt ttgtggctgt gcagaacttg
     1081 gaggactggg aggaagccat cttcgacaag agcacaggga aggttgtttt gaagacgttc
     1141 agcctctaca agaagctgct gactcttttc agagctggcc acgaccaggt ggtggtcctg
     1201 ctccatgatg tccgtgatgt gagcgtggag gaggagaagg tccggtactt cgggaaaggc
     1261 tacatggtgg tgctccggct tgcgacgggc ttctcccacc ccctcacgca gagtgcagtc
     1321 atgggccacc gcagtgatgt ggaagccatc gccaagctca tcaccagctt cctggagctg
     1381 cactgccttg agagccccac agagctgtct cagagcagcg acagtgaggc cggtgaccct
     1441 gcaagccaga gcgtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc
     1501 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1561 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     1621 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1681 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1741 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     1801 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     1861 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1921 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     1981 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2041 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2101 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2161 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2221 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2281 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2341 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     2401 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     2461 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2521 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2581 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     2641 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     2701 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     2761 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     2821 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     2881 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     2941 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3001 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3061 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3121 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3181 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3241 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     3301 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     3361 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     3421 gggtcagcac cgtttctgcg gactggcttt ctacccggta tcagctcact caaaggcggt
     3481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4141 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     4201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4261 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     4321 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     4381 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     4441 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     4501 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     4561 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     4621 tct
//
