LOCUS       sequence-471844-        5730 bp    DNA     circular SYN 09-MAY-2025
DEFINITION  Express ARRB1-SmBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5730)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: ARRB1-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2619..2740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2630..2649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2855..2888)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2898..2945
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2977..3771
                     /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(3031..3050)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3641..3660
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3988..4533
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4434..4453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4650..4667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4817..4865
                     /note="synthetic polyadenylation signal"
     CDS             complement(4889..5287)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5284..5293
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5318..5675)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5332..5467
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5484..5675
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5632..5652
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat gggcgacaaa
     1201 gggacccgag tgttcaagaa ggccagtcca aatggaaagc tcaccgtcta cctgggaaag
     1261 cgggactttg tggaccacat cgacctcgtg gaccctgtgg atggtgtggt cctggtggat
     1321 cctgagtatc tcaaagagcg gagagtctat gtgacgctga cctgcgcctt ccgctatggc
     1381 cgggaggacc tggatgtcct gggcctgacc tttcgcaagg acctgtttgt ggccaacgta
     1441 cagtcgttcc caccggcccc cgaggacaag aagcccctga cgcggctgca ggaacgcctc
     1501 atcaagaagc tgggcgagca cgcttaccct ttcacctttg agatccctcc aaaccttcca
     1561 tgttctgtga cactgcagcc ggggcccgaa gacacgggga aggcttgcgg tgtggactat
     1621 gaagtcaaag ccttctgcgc ggagaatttg gaggagaaga tccacaagcg gaattctgtg
     1681 cgtctggtca tccggaaggt tcagtatgcc ccagagaggc ctggccccca gcccacagcc
     1741 gagaccacca ggcagttcct catgtcggac aagcccttgc acctagaagc ctctctggat
     1801 aaggagatct attaccatgg agaacccatc agcgtcaacg tccacgtcac caacaacacc
     1861 aacaagacgg tgaagaagat caagatctca gtgcgccagt atgcagacat ctgccttttc
     1921 aacacagctc agtacaagtg ccctgttgcc atggaagagg ctgatgacac tgtggcaccc
     1981 agctcgacgt tctgcaaggt ctacacactg acccccttcc tagccaataa ccgagagaag
     2041 cggggcctcg ccttggacgg gaagctcaag cacgaagaca cgaacttggc ctctagcacc
     2101 ctgttgaggg aaggtgccaa ccgtgagatc ctggggatca ttgtttccta caaagtgaaa
     2161 gtgaagctgg tggtgtctcg gggcggcctg ttgggagatc ttgcatccag cgacgtggcc
     2221 gtggaactgc ccttcaccct aatgcacccc aagcccaaag aggaaccccc gcatcgggaa
     2281 gttccagaga acgagacgcc agtagatacc aatctcatag aacttgacac aaatgatgac
     2341 gacattgtat ttgaggactt tgctcgccag agactgaaag gcatgaagga tgacaaggag
     2401 gaagaggagg atggtaccgg ctctccacag ctcaacaaca gagtttctca aggcagttca
     2461 ggtggtggcg ggagcggagg tggaggctcg agcggtgtga ccggctaccg gctgttcgag
     2521 gagattctgt aaggccgcga ctctagagtc gacctgcagg catgcaagct gatttggctg
     2581 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2761 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     2821 caaatgtggt aaaatcttaa ttaagaattc taatataact tcgtatagca tacattatac
     2881 gaagttatgg atccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     2941 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     3001 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3061 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3121 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3181 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3241 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3301 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3361 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3421 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3481 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3541 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3601 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3661 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3721 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     3781 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     3841 cagaatcagg ggataacgca ggaaagaaca tgtcaaacat gagaattaca acttatatcg
     3901 tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa tctagaaata
     3961 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
     4021 ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa
     4081 ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt
     4141 ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg
     4201 ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata
     4261 aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg
     4321 cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg
     4381 acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac
     4441 gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg
     4501 tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt
     4561 ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt
     4621 ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata
     4681 tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag
     4741 cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc
     4801 tgatgtccgg aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt
     4861 ttattgcggc catcgtgatg gctagagttt agccctccca cacataacca gagggcagca
     4921 attcacgaat cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc
     4981 caggatgcag atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt
     5041 tctcatttcc gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc
     5101 ccagcaccac gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga
     5161 agatgcggcc gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga
     5221 tgctgttgat tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct
     5281 tggccatggt ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa
     5341 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     5401 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     5461 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     5521 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     5581 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     5641 gacacacatt ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt
     5701 cagaggttat ttcaggccat ggtgctgcgc
//
