LOCUS       sequence-450120-        4053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid can be used to monitor receptor internalization in real
            time living cells using bioluminescence as a readout..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4053)
  AUTHORS   Reyes-Alcaraz A, Lucero Garcia-Rojas EY, Merlinsky EA, Seong JY, 
            Bond RA, McConnell BK
  TITLE     A NanoBiT assay to monitor membrane proteins trafficking for drug 
            discovery and drug development.
  JOURNAL   Commun Biol. 2022 Mar 8;5(1):212. doi: 10.1038/s42003-022-03163-9.
  PUBMED    35260793
REFERENCE   2  (bases 1 to 4053)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2022-03-8"; volume: "5(1):212. doi"; pages: " 
            10.1038/s42003-022-03163-9"
COMMENT     Sequence Label: MCS-3_Endofin_LgBiT
FEATURES             Location/Qualifiers
     source          1..4053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..779
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      809..818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1607..1728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1618..1637)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1672..1691
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1976..1993)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2147..2735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2227..2246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2935..3726,3727..3795))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3900..3948
                     /note="synthetic polyadenylation signal"
     misc_feature    3962..4053
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4002..4021
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagtctaaat gagtcttcgg acctcgcggg ggccgcttaa
       61 gcggtggtta gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg
      121 gaggcggctc ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc
      181 ttaagcaccc ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg
      241 gccgctgact cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac
      301 ccccgggcgg cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc
      361 gtatacgtgg acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc
      421 tgcgggacga gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg
      481 acggggcaca ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg
      541 gagtggcgca gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg
      601 gaagaaatat atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg
      661 tcattggcga agtcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg
      721 catattaagg tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa
      781 agcttggcaa tccggtactg ttggtaaagc caccatggtc ttcacactcg aagatttcgt
      841 tggggactgg gaacagacag ccgcctacaa cctggaccaa gtccttgaac agggaggtgt
      901 gtccagtttg ctgcagaatc tcgccgtgtc cgtaactccg atccaaagga ttgtccggag
      961 cggtgaaaat gccctgaaga tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgc
     1021 cgaccaaatg gcccagatcg aagaggtgtt taaggtggtg taccctgtgg atgatcatca
     1081 ctttaaggtg atcctgccct atggcacact ggtaatcgac ggggttacgc cgaacatgct
     1141 gaactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa agatcactgt
     1201 aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca cccccgacgg
     1261 ctccatgctg ttccgagtaa ccatcaacag tgggagttcc ggtggtggcg ggagcggagg
     1321 tggaggctcg agcggtggag ctcaggggaa ttcacagaaa cagcctactt gggttcctga
     1381 ttcagaagct ccaaactgta tgaactgcca agtcaaattt acttttacca aacggcgaca
     1441 ccattgccga gcatgtggga aagtattttg tggtgtctgt tgtaatagga agtgtaaact
     1501 gcaatatcta gaaaaggaag caagagtatg tgtagtctgc tatgaaacta ttagtaaata
     1561 aagatcttct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     1621 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1681 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1741 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1801 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     1861 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     1921 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     1981 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2041 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2101 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2161 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2221 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2281 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2341 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2401 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2461 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2521 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2581 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2641 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2701 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2821 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2881 tatatgagta aacttggtct gacagcggcc gcaaatgcta aaccactgca gtggttacca
     2941 gtgcttgatc agtgaggcac cgatctcagc gatctgccta tttcgttcgt ccatagtggc
     3001 ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc ttaccatcag gccccagcgc
     3061 agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat ttgtcagcaa tgaaccagcc
     3121 agcagggagg gccgagcgaa gaagtggtcc tgctactttg tccgcctcca tccagtctat
     3181 gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg agtagtttcc gaagagttgt
     3241 ggccattgct actggcatcg tggtatcacg ctcgtcgttc ggtatggctt cgttcaactc
     3301 tggttcccag cggtcaagcc gggtcacatg atcacccata ttatgaagaa atgcagtcag
     3361 ctccttaggg cctccgatcg ttgtcagaag taagttggcc gcggtgttgt cgctcatggt
     3421 aatggcagca ctacacaatt ctcttaccgt catgccatcc gtaagatgct tttccgtgac
     3481 cggcgagtac tcaaccaagt cgttttgtga gtagtgtata cggcgaccaa gctgctcttg
     3541 cccggcgtct atacgggaca acaccgcgcc acatagcagt actttgaaag tgctcatcat
     3601 cgggaatcgt tcttcggggc ggaaagactc aaggatcttg ccgctattga gatccagttc
     3661 gatatagccc actcttgcac ccagttgatc ttcagcatct tttactttca ccagcgtttc
     3721 ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag ggaatgagtg cgacacgaaa
     3781 atgttggatg ctcatactcg tcctttttca atattattga agcatttatc agggttacta
     3841 gtacgtctct caaggataag taagtaatat taaggtacgg gaggtattgg acaggccgca
     3901 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     3961 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     4021 ccagtgcaag tgcaggtgcc agaacatttc tct
//
