LOCUS       sequence-471843-        5789 bp    DNA     circular SYN 09-MAY-2025
DEFINITION  Express SmBiT-ARRB1 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5789)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5789)
  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: SmBiT-ARRB1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5789
                     /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    2678..2799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2689..2708)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2914..2947)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2957..3004
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3036..3830
                     /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(3090..3109)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3700..3719
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4047..4592
                     /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     4493..4512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4709..4726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4876..4924
                     /note="synthetic polyadenylation signal"
     CDS             complement(4948..5346)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5343..5352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5377..5734)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5391..5526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5543..5734
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5691..5711
                     /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 cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat gggcgacaaa gggacccgag tgttcaagaa ggccagtcca
     1321 aatggaaagc tcaccgtcta cctgggaaag cgggactttg tggaccacat cgacctcgtg
     1381 gaccctgtgg atggtgtggt cctggtggat cctgagtatc tcaaagagcg gagagtctat
     1441 gtgacgctga cctgcgcctt ccgctatggc cgggaggacc tggatgtcct gggcctgacc
     1501 tttcgcaagg acctgtttgt ggccaacgta cagtcgttcc caccggcccc cgaggacaag
     1561 aagcccctga cgcggctgca ggaacgcctc atcaagaagc tgggcgagca cgcttaccct
     1621 ttcacctttg agatccctcc aaaccttcca tgttctgtga cactgcagcc ggggcccgaa
     1681 gacacgggga aggcttgcgg tgtggactat gaagtcaaag ccttctgcgc ggagaatttg
     1741 gaggagaaga tccacaagcg gaattctgtg cgtctggtca tccggaaggt tcagtatgcc
     1801 ccagagaggc ctggccccca gcccacagcc gagaccacca ggcagttcct catgtcggac
     1861 aagcccttgc acctagaagc ctctctggat aaggagatct attaccatgg agaacccatc
     1921 agcgtcaacg tccacgtcac caacaacacc aacaagacgg tgaagaagat caagatctca
     1981 gtgcgccagt atgcagacat ctgccttttc aacacagctc agtacaagtg ccctgttgcc
     2041 atggaagagg ctgatgacac tgtggcaccc agctcgacgt tctgcaaggt ctacacactg
     2101 acccccttcc tagccaataa ccgagagaag cggggcctcg ccttggacgg gaagctcaag
     2161 cacgaagaca cgaacttggc ctctagcacc ctgttgaggg aaggtgccaa ccgtgagatc
     2221 ctggggatca ttgtttccta caaagtgaaa gtgaagctgg tggtgtctcg gggcggcctg
     2281 ttgggagatc ttgcatccag cgacgtggcc gtggaactgc ccttcaccct aatgcacccc
     2341 aagcccaaag aggaaccccc gcatcgggaa gttccagaga acgagacgcc agtagatacc
     2401 aatctcatag aacttgacac aaatgatgac gacattgtat ttgaggactt tgctcgccag
     2461 agactgaaag gcatgaagga tgacaaggag gaagaggagg atggtaccgg ctctccacag
     2521 ctcaacaaca gagtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc
     2581 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     2641 tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2701 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2761 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2821 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac
     2881 aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat acattatacg
     2941 aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca tagtataata
     3001 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     3061 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3121 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3181 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3241 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3301 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3361 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3421 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3481 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3541 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3601 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3661 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3721 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3781 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3841 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3901 agaatcaggg gataacgcag gaaagaacat gtcaaacatg agaattacaa cttatatcgt
     3961 atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat
     4021 tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     4081 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     4141 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     4201 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     4261 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     4321 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     4381 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     4441 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     4501 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt
     4561 gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc
     4621 cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc
     4681 cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat
     4741 cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc
     4801 acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct
     4861 gatgtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     4921 tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag agggcagcaa
     4981 ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc
     5041 aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt
     5101 ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc
     5161 cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa
     5221 gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat
     5281 gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt
     5341 ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     5401 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     5461 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     5521 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     5581 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     5641 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     5701 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     5761 agaggttatt tcaggccatg gtgctgcgc
//
