LOCUS       sequence-308909-        5701 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expresses squirrel monkey CHMP3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  AUTHORS   Rheinemann L, Downhour DM, Bredbenner K, Mercenne G, Davenport KA, 
            Schmitt PT, Necessary CR, McCullough J, Schmitt AP, Simon SM, 
            Sundquist WI, Elde NC
  TITLE     RetroCHMP3 blocks budding of enveloped viruses without blocking 
            cytokinesis.
  JOURNAL   Cell. 2021 Sep 27. pii: S0092-8674(21)01055-2. doi: 
            10.1016/j.cell.2021.09.008.
  PUBMED    34597582
REFERENCE   2  (bases 1 to 5701)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2021 
            Sep 27. pii: S0092-8674(21)01055-2. doi: 
            10.1016/j.cell.2021.09.008."
COMMENT     Sequence Label: pCMV(Delta-2)-squirrel-monkey-full-length-CHMP3-HA
FEATURES             Location/Qualifiers
     source          1..5701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      150..578
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(237..256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     447..468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(587..607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     801..820
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             955..1749
                     /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(1009..1028)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1619..1638
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1923..2044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1960..1979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2014..2033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2093..2109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2106..2128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2117..2133
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2141..2147,2148..2165,2166..2171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2186..2207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2324..2341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2495..3083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2575..2594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3254..4045,4046..4114))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3877..3896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4115..4219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4294..4313)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4552..4651
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     4602..4622
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4696..4715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4696..4714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5456..5482
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5501..5520)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     complement(5575..5592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(5581..5701,1..104)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
       61 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
      121 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
      181 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      241 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      301 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
      361 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
      421 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
      481 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
      541 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
      601 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      661 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      721 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
      781 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
      841 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcgaaaa gctcccggga
      901 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
      961 gaacaagatg gattgcacgc aggttctccg gctgcttggg tggagaggct attcggctat
     1021 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     1081 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     1141 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     1201 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     1261 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     1321 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     1381 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     1441 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     1501 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     1561 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     1621 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     1681 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     1741 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     1801 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     1861 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     1921 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2041 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     2101 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2161 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2221 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2281 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3001 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3061 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3121 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3181 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3241 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3301 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3361 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3421 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3481 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3541 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3601 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3661 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3721 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3781 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3841 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3901 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3961 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4021 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4081 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4141 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4201 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     4261 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4321 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     4381 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     4441 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttagcggt ttgactcacg
     4501 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     4561 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     4621 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     4681 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agcgtttaaa
     4741 cgggcccggg aacggaaagc cggaaggagc aagacgagtt cagttcgcca tggggctgtt
     4801 tggaaaaacc caggagaagc cacccaaaga actggtcaat gaatggtcat tgaagataag
     4861 aaaggaaatg agagttgttg acaggcaaat aagggatatc caaagagaag aagaaaaagt
     4921 gaaacgatct gtgaaagatg cagccaagaa gggccagaag gatgtctgtg tagttctggc
     4981 caaggagatg atcaggtcaa ggaaggccgt gagcaagcta tatgcatcca aagcgcacat
     5041 gaattcggtg ctcatgggga tgaagaacca gctcgcggtc ttgcgagtgg ctggttccct
     5101 gcagaagagc acagaagtga tgaaggccat gcaaagtctt gtgaagattc cagaaattca
     5161 ggccaccatg agggagctat ccaaagaaat gatgaaggct gggatcatag aggagatgtt
     5221 agaggacact tttgaaagca tggacgatca ggaagaaatg gaggaagaag cagaaatgga
     5281 aattgacaaa attctatttg aaattacagc aggggccttg ggcaaagcac ccagtaaagt
     5341 gactgatgcc cttccagagc cagaaccttc gggagcgatg gctgcctcag aggatgagga
     5401 ggaggaggaa gaagctctgg aggccatgca gtcccggctg gccacactcc gaagctaccc
     5461 atacgatgtt ccagattacg cttgaaagct tgctagcaga tctttttccc tctgccaaaa
     5521 attatgggga catcatgaag ccccttgagc acttaagttt aaaccgctga tcagcctcga
     5581 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5641 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5701 t
//
