LOCUS       sequence-472396-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SmBiT-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 5552)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5552)
  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            "Unpublished"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: SmBiT-ARRB2 Vector (CMV)
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        198..577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        578..781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     732..752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          917..1049
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1032..1051
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1093..1112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1093..1111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2555..2573)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2610..2731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2621..2640)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2675..2694
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2976..3333
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2976..3167
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2999..3019)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3184..3319
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3348..3395
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3427..4221
                     /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(3481..3500)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4091..4110
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4285..4333
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4572..5160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4652..4671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(5276..5552,1..7)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
       61 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
      121 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      181 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      421 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      721 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      781 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      841 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      901 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      961 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1021 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1081 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg accggctacc
     1141 ggctgttcga ggagattctc gggagctccg gtggtggcgg gagcggaggt ggaggctcga
     1201 gcggtgcgat cgccatgggg gagaaacccg ggaccagggt cttcaagaag tcgagcccta
     1261 actgcaagct caccgtgtac ttgggcaagc gggacttcgt agatcacctg gacaaagtgg
     1321 accctgtaga tggcgtggtg cttgtggacc ctgactacct gaaggaccgc aaagtgtttg
     1381 tgaccctcac ctgcgccttc cgctatggcc gtgaagacct ggatgtgctg ggcttgtcct
     1441 tccgcaaaga cctgttcatc gccacctacc aggccttccc cccggtgccc aacccacccc
     1501 ggccccccac ccgcctgcag gaccggctgc tgaggaagct gggccagcat gcccacccct
     1561 tcttcttcac cataccccag aatcttccat gctccgtcac actgcagcca ggcccagagg
     1621 atacaggaaa ggcctgcggc gtagactttg agattcgagc cttctgtgct aaatcactag
     1681 aagagaaaag ccacaaaagg aactctgtgc ggctggtgat ccgaaaggtg cagttcgccc
     1741 cggagaaacc cggcccccag ccttcagccg aaaccacacg ccacttcctc atgtctgacc
     1801 ggtccctgca cctcgaggct tccctggaca aggagctgta ctaccatggg gagcccctca
     1861 atgtaaatgt ccacgtcacc aacaactcca ccaagaccgt caagaagatc aaagtctctg
     1921 tgagacagta cgccgacatc tgcctcttca gcaccgccca gtacaagtgt cctgtggctc
     1981 aactcgaaca agatgaccag gtatctccca gctccacatt ctgtaaggtg tacaccataa
     2041 ccccactgct cagcgacaac cgggagaagc ggggtctcgc cctggatggg aaactcaagc
     2101 acgaggacac caacctggct tccagcacca tcgtgaagga gggtgccaac aaggaggtgc
     2161 tgggaatcct ggtgtcctac agggtcaagg tgaagctggt ggtgtctcga ggcggggatg
     2221 tctctgtgga gctgcctttt gttcttatgc accccaagcc ccacgaccac atccccctcc
     2281 ccagacccca gtcagccgct ccggagacag atgtccctgt ggacaccaac ctcattgaat
     2341 ttgataccaa ctatgccaca gatgatgaca ttgtgtttga ggactttgcc cggcttcggc
     2401 tgaaggggat gaaggatgac gactatgatg atcaactctg cgtttaaacg aattcgggct
     2461 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagctgatcc ggctgctaac
     2521 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2581 cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2641 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2701 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2761 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     2821 taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     2881 tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa
     2941 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     3001 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3061 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3181 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3241 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3301 ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt gacaattaat
     3361 catcggcata gaatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg
     3421 cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3481 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3541 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3601 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3661 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3721 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3781 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3841 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3901 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3961 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4021 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4081 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4141 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4201 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4261 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     4321 ggttttttgt gtgaatcgat agcgataagg atcctctttg cgcttgcgtt ttcccttgtc
     4381 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     4441 tacccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4741 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4921 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4981 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5041 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5101 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
     5161 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5221 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
     5281 aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca
     5341 gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat
     5401 acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt
     5461 acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt
     5521 atggtaaatc cacttactgt ctgccctcgt ag
//
