LOCUS       sequence-274403-        4794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human CHMP4B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4794)
  TITLE     Mammalian expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4794)
  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            "Unpublished"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: HA-CHMP4B
FEATURES             Location/Qualifiers
     source          1..4794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1070..1096
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1800..1819)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1801..1819)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1824..1958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1848..1867)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1902..1921
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2071..2091)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2071..2089)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2110..2126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2110..2126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2123..2145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2134..2150
                     /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(2158..2164,2165..2182,2183..2188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2203..2224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2341..2358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2512..3100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2592..2611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3271..4062,4063..4131))
                     /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     3894..3913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4132..4236)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4262..4717)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4399..4420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4611..4630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat accatgcgat acccatatga
     1081 tgtgccagat tacgctggat ctatgtcggt gttcgggaag ctgttcgggg ctggaggggg
     1141 taaggccggc aagggcggcc cgacccccca ggaggccatc cagcggctgc gggacacgga
     1201 agagatgtta agcaagaaac aggagttcct ggagaagaaa atcgagcagg agctgacggc
     1261 cgccaagaag cacggcacca aaaacaagcg cgcggccctc caggcactga agcgtaagaa
     1321 gaggtatgag aagcagctgg cgcagatcga cggcacatta tcaaccatcg agttccagcg
     1381 ggaggccctg gagaatgcca acaccaacac cgaggtgctc aagaacatgg gctatgccgc
     1441 caaggccatg aaggcggccc atgacaacat ggacatcgat aaagttgatg agttaatgca
     1501 ggacattgct gaccagcaag aacttgcaga ggagatttca acagcaattt cgaaacctgt
     1561 agggtttgga gaagagtttg acgaggatga gctcatggcg gaattagaag aactagaaca
     1621 ggaggaacta gacaagaatt tgctggaaat cagtggaccc gaaacagtcc ctctaccaaa
     1681 tgttccctct atagccctac catcaaaacc cgccaagaag aaagaagagg aggacgacga
     1741 catgaaggaa ttggagaact gggctggatc catgtaactc gagcctctag attctgcagc
     1801 cctatagtga gtcgtattac gtagatccag acatgataag atacattgat gagtttggac
     1861 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1921 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1981 ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa
     2041 tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     2101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2161 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2941 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4261 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4321 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4381 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4441 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4501 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4561 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4621 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4681 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     4741 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gacc
//
