LOCUS       sequence-472500-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express LgBiT-ARRB2 Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5993)
  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: 
            "Unpublished"
COMMENT     Sequence Label: LgBiT-ARRB2 Vector
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(778..797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(779..797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(839..858)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          841..973
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(1109..1312)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1138..1158)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1313..1692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1883..2166
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      2282..2870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2771..2790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3109..3157
                     /note="synthetic polyadenylation signal"
     CDS             complement(3221..4015)
                     /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(3332..3351)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3942..3961
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4047..4094)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4109..4466)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4123..4258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4275..4466
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4423..4443
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4711..4832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4748..4767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4802..4821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4869..4887
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 tttgcggaag gacaagccca gcacatccag gtcttcacgg ccatagcgga aggcgcaggt
       61 gagggtcaca aacactttgc ggtccttcag gtagtcaggg tccacaagca ccacgccatc
      121 tacagggtcc actttgtcca ggtgatctac gaagtcccgc ttgcccaagt acacggtgag
      181 cttgcagtta gggctcgact tcttgaagac cctggtcccg ggtttctccc ccatggcgat
      241 cgcaccgctc gagcctccac ctccgctccc gccaccaccg gagctcccac tgttgatggt
      301 tactcggaac agcatggagc cgtcgggggt gatcaggcgc tcgtcgataa ttttgttgcc
      361 gttccacagg gtccctgtta cagtgatctt tttgccgtcg aacacggcga tgccttcata
      421 cggccgtccg aaatagttca gcatgttcgg cgtaaccccg tcgattacca gtgtgccata
      481 gggcaggatc accttaaagt gatgatcatc cacagggtac accaccttaa acacctcttc
      541 gatctgggcc atttggtcgg cgctcagacc ttcatacggg atgatgacat ggatgtcgat
      601 cttcagggca ttttcaccgc tccggacaat cctttggatc ggagttacgg acacggcgag
      661 attctgcagc aaactggaca cacctccctg ttcaaggact tggtccaggt tgtaggcggc
      721 tgtctgttcc cagtccccaa cgaaatcttc gagtgtgaag accatggtga gcgctagccc
      781 tatagtgagt cgtattaata ctctagcctt aagagctgta attgaactgg gagtggacac
      841 ctgtggagag aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca
      901 agagtcttct ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa
      961 ccttgatact tacctgccca gtgcctcacg accaacttct gcagcttaag ttcgagactg
     1021 ttgtgtcaga agcactgact gcgttagcaa tttaactgtg ataaactacc gcaataaagc
     1081 ttctagtgat ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac
     1141 gcctaccgcc catttgcgtc aacggggcgg ggttattacg acattttgga aagtcccgtt
     1201 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     1261 gtgagtcaaa ccgctatcca cgcccattgg tgtactgcca aaaccgcatc accatggtaa
     1321 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccgtaag gtcatgtact
     1381 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc ggacttggca
     1441 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     1501 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     1561 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     1621 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     1681 ataatcaatg ccaacatggc ggtcatattg gacatgagcc aatataaatg tacatattat
     1741 gatatagata caacgtatgc aatggccaat agccaatatt gatttatgct atataaccaa
     1801 tgaataatat ggctaatggc caatattgaa gatctcgatc ctctacgccg gacgcatcgt
     1861 ggccggagta ctgcggttta tctcgatggc tacgagggca gacagtaagt ggatttacca
     1921 taatccctta attgtacgca ccgctaaaac gcgttcagcg cgatcacggc agcagacagg
     1981 taaaaatggc aacaaaccac ccgaaaaact gccgcaatcg cgcctgataa attttaaccg
     2041 tatgaatacc tatgcaacca gagggtacag gccacattac ccccacttaa tccactgaag
     2101 ctgccatttt tcatggtttc accatcccag cgaagggcca tccagcgtgc gttcctgtat
     2161 ttccggacta taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2221 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2281 cttgaaatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2341 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2401 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2461 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2521 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2581 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2641 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2701 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2761 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2821 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2881 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2941 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgggt
     3001 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     3061 ggacaaggga aaacgcaagc gcaaagagga tccttatcgc tatcgattca cacaaaaaac
     3121 caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga tggcaggttg
     3181 ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     3241 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     3301 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     3361 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     3421 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     3481 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     3541 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     3601 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     3661 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     3721 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     3781 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     3841 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     3901 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     3961 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgatct
     4021 gcctcctggg tttagttcct caccttgtcg tattatacta tgccgatata ctatgccgat
     4081 gattaattgt caacagttaa ttaagctttt tgcaaaagcc taggcctcca aaaaagcctc
     4141 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     4201 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     4261 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     4321 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     4381 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     4441 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     4501 tttcaggcca tggtgctgcg cagatccgcg tatgcggtgt gaaataccgc acagatgcgt
     4561 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgatttt
     4621 accacatttg tagaggtttt acttgcttaa aaaaacctcc cacatctccc cctgaacctg
     4681 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4741 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4801 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gctcgaagcg gccgccccaa
     4861 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
     4921 tgttagcagc cggatcagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc
     4981 gagcccgaat tcgtttaaac gcagagttga tcatcatagt cgtcatcctt catccccttc
     5041 agccgaagcc gggcaaagtc ctcaaacaca atgtcatcat ctgtggcata gttggtatca
     5101 aattcaatga ggttggtgtc cacagggaca tctgtctccg gagcggctga ctggggtctg
     5161 gggaggggga tgtggtcgtg gggcttgggg tgcataagaa caaaaggcag ctccacagag
     5221 acatccccgc ctcgagacac caccagcttc accttgaccc tgtaggacac caggattccc
     5281 agcacctcct tgttggcacc ctccttcacg atggtgctgg aagccaggtt ggtgtcctcg
     5341 tgcttgagtt tcccatccag ggcgagaccc cgcttctccc ggttgtcgct gagcagtggg
     5401 gttatggtgt acaccttaca gaatgtggag ctgggagata cctggtcatc ttgttcgagt
     5461 tgagccacag gacacttgta ctgggcggtg ctgaagaggc agatgtcggc gtactgtctc
     5521 acagagactt tgatcttctt gacggtcttg gtggagttgt tggtgacgtg gacatttaca
     5581 ttgaggggct ccccatggta gtacagctcc ttgtccaggg aagcctcgag gtgcagggac
     5641 cggtcagaca tgaggaagtg gcgtgtggtt tcggctgaag gctgggggcc gggtttctcc
     5701 ggggcgaact gcacctttcg gatcaccagc cgcacagagt tccttttgtg gcttttctct
     5761 tctagtgatt tagcacagaa ggctcgaatc tcaaagtcta cgccgcaggc ctttcctgta
     5821 tcctctgggc ctggctgcag tgtgacggag catggaagat tctggggtat ggtgaagaag
     5881 aaggggtggg catgctggcc cagcttcctc agcagccggt cctgcaggcg ggtggggggc
     5941 cggggtgggt tgggcaccgg ggggaaggcc tggtaggtgg cgatgaacag gtc
//
