LOCUS       sequence-479826-        5064 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express LgBiT-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 5064)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5064)
  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            "Unpublished"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: LgBiT-CYBC1 (TK) Vector
FEATURES             Location/Qualifiers
     source          1..5064
                     /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(2007..2025)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2062..2183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2073..2092)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2127..2146
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2428..2785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2428..2619
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2451..2471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2636..2771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2800..2847
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2879..3673
                     /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(2933..2952)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3543..3562
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3737..3785
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4024..4612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4104..4123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4728..5011
                     /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 catggtcttc acactcgaag atttcgttgg ggactgggaa
      841 cagacagccg cctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgctg
      901 cagaatctcg ccgtgtccgt aactccgatc caaaggattg tccggagcgg tgaaaatgcc
      961 ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcgccga ccaaatggcc
     1021 cagatcgaag aggtgtttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc
     1081 ctgccctatg gcacactggt aatcgacggg gttacgccga acatgctgaa ctatttcgga
     1141 cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg
     1201 tggaacggca acaaaattat cgacgagcgc ctgatcaccc ccgacggctc catgctgttc
     1261 cgagtaacca tcaacagtgg gagctccggt ggtggcggga gcggaggtgg aggctcgagc
     1321 ggtgcgatcg ccatgtacct gcaggtggag acccgcacca gctcccgcct ccatctgaag
     1381 agggctccag gcatccggtc ctggtccctg ctggttggaa tcttgtcgat tggcctggct
     1441 gctgcctact acagcggaga tagcctgggc tggaagctct tctacgtcac aggctgcctg
     1501 tttgtggctg tgcagaactt ggaggactgg gaggaagcca tcttcgacaa gagcacaggg
     1561 aaggttgttt tgaagacgtt cagcctctac aagaagctgc tgactctttt cagagctggc
     1621 cacgaccagg tggtggtcct gctccatgat gtccgtgatg tgagcgtgga ggaggagaag
     1681 gtccggtact tcgggaaagg ctacatggtg gtgctccggc ttgcgacggg cttctcccac
     1741 cccctcacgc agagtgcagt catgggccac cgcagtgatg tggaagccat cgccaagctc
     1801 atcaccagct tcctggagct gcactgcctt gagagcccca cagagctgtc tcagagcagc
     1861 gacagtgagg ccggtgaccc tgcaagccag agcgtttaaa cgaattcggg ctcggtaccc
     1921 ggggatcctc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
     1981 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     2041 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2101 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2161 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2221 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     2281 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     2341 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     2401 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     2461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2701 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2761 ttttggaggc ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca
     2821 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     2881 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2941 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3001 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3061 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3121 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3181 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3241 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3301 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3361 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3421 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3481 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3541 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3601 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3661 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3721 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     3781 gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc
     3841 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt
     3901 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3961 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4441 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     4621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     4741 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     4801 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     4861 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     4921 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     4981 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     5041 gtccggcgta gaggatcgag atct
//
