LOCUS       sequence-310422-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally HA-tagged PfROM4 variant H578A (codon 
            optimized for mammalian expression) from pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  AUTHORS   Gandhi S, Baker RP, Cho S, Stanchev S, Strisovsky K, Urban S
  TITLE     Designed Parasite-Selective Rhomboid Inhibitors Block Invasion and 
            Clear Blood-Stage Malaria.
  JOURNAL   Cell Chem Biol. 2020 Aug 31. pii: S2451-9456(20)30333-0. doi: 
            10.1016/j.chembiol.2020.08.011.
  PUBMED    32888502
REFERENCE   2  (bases 1 to 7795)
  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: "Cell Chem 
            Biol. 2020 Aug 31. pii: S2451-9456(20)30333-0. doi: 
            10.1016/j.chembiol.2020.08.011."
COMMENT     Sequence Label: pcDNA-HA-PfROM4 (H578A)
FEATURES             Location/Qualifiers
     source          1..7795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             924..1013
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(3386..3403)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3392..3616
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3662..4090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3749..3768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3959..3980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4099..4119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4104..4433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4284..4419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4346..4365
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4500..5294
                     /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(4554..4573)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5164..5183
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5468..5589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5505..5524)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5559..5578
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5638..5654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5638..5654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5651..5673)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5662..5678
                     /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(5686..5692,5693..5710,5711..5716))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5731..5752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5869..5886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6040..6628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6120..6139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6799..7590,7591..7659))
                     /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     7422..7441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7660..7764)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga atgtacccat acgatgttcc tgactatgcg ggctatccct
      961 atgacgtccc ggactatgca ggatcctatc catatgacgt tccagattac gctggcgaat
     1021 tcggcagcaa taccggcttc aaccccaaga acgccgagaa gaagcggagc agcatcttca
     1081 gcgacctgga gatcccccag ggcgacgtga agaagaaggc ctccatcttc aataatatgg
     1141 agaagaaaaa gatgaccatc ttcaagaaaa ccaagggcga gaacaacgag gagaagaaga
     1201 aaagaagcat cttcaataac aataataaca acaataataa taacaacacc atgaagatca
     1261 acatctttaa caataacaac gagaagaaag aaagcttctc cattaacggg gatgaaaaga
     1321 agaaatcctt ttccataaat ggcgacgaga aaaaggagtc tttttccatt aacggagatg
     1381 aaaaaaagga gagcttcctg atcaacaaag acgagaaaaa agagtctttc ctgattaata
     1441 aggatgaaaa ggagaagagc ttctctatca atggcgacga aaagaaagag agcttttcta
     1501 ttaacgggga tgaaaagaag gaatccttta gcatcaatga ggacgagacc aagaagagct
     1561 gttccatcaa cgacgatgag cggaagatct ccatcttctc caataacgaa aagaagaaga
     1621 acagcatcta ctccgacacc cagagcagca agcgggagga tgagaagcgg atctccattt
     1681 ttagcgatct ggagaccagc accaacatcg acgacatcag ctccaagcgg aacagccaga
     1741 agctgtccgt gtacggcgac aacaagctga agaagggcag cctgctgtcc cctaaggtgg
     1801 acaactaccg gaacaccatc gacaacatca acgagatcaa cgacatcaag atcatcgtga
     1861 ccagcgacga gaatctgcac accctgccca gcggagctgt gggcagaaga gcccccctga
     1921 accctttcag cagccccatc ctgggcaagt accggcggaa gaacaagaac gccaaggcca
     1981 aggtgaagga cccccggctg aacaacaatc ccctgatcgg caggctgacc gtgtgtatca
     2041 gcaccaccgc catcctgttc tgggtgttct tcgccgagat ggtgttcaac tacaacacct
     2101 tcaacggccg gtgtatctcc aaggtgctgt accccatcta caccgagaac gtggtgctga
     2161 aacggcagcc cttcttcgtg ttcctgggct acggcgcctg tgagtacaac ctggacgaga
     2221 gcgccagcaa cagacacttc atcggcagca aggccagcga tgagggctgg cctggcgaca
     2281 aggtggagga gaaccctgat ggcagaggct acgccaactg ggacagcgtg aacaaccggg
     2341 tgtacaatct gctgggcggc ctgaacacca actacatccg gaactacggc gagctgtaca
     2401 ggctgttctg gagcatgtac ctgcacggcg gcttcatgca catcctgttc aacgtgatct
     2461 gccagatcca gatcctgtgg atgatcgagc ctgactgggg cagcatcaga accggcctgc
     2521 tgttctttat tagcggcgtg accggcaatc tgctgtccgc cgtgtgtgac ccctgtggcg
     2581 tgaccatcgg cagctccggc agcctgtacg gcctgatcgg cgccctgttc gcctactaca
     2641 tcgagtactg gaaaaccatc cccaggccct gctgtgtgct gatcttcatg tttctggtgg
     2701 tgatgttcgg catcattgtg ggcatgttcg gctacaccga caactacgcc gccatcggcg
     2761 gctgcctggg cggcgtgctg ttcggattca gcacaatcac caccgtgtct gccgccgata
     2821 agtgtaccct gggcgagaga atgctggtgt ccgccccctt cagctggttc ctgagcaatg
     2881 agaccaagga gctgatcatc gccaaagcca aggacaagaa gatcaagggg gagaacttcc
     2941 ggaagaagca gctggccaac aaggtgcaca agaacgacgc cctgcacgtg gccatggctg
     3001 tcatgaagaa ccggatcaac gatgagggca gacctccttg ccggatgaag ctgagggagt
     3061 ggattgtgag aatcaccgcc gccagcaccc tgatcatcat gtggatcgtg ctgttcatct
     3121 acctgctgaa cgagaaggcc tacaagagct acagccccct gggccagatc aagtttagcg
     3181 gcgtgcacag ctgctactgc tgccagatcg tgaagaacaa gttcacctac atcaaggtga
     3241 acgacttcta ctggtgcttc accaccgagg aggccacccg gtactactgt aacaagtgat
     3301 agtctagaac gcggagctcc agcttttgtt ccctttagtg agcggccgca gcaggtacca
     3361 agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3421 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3481 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3541 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3601 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3661 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3721 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3781 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3841 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3901 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3961 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4021 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4081 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4141 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4201 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4261 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4321 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4381 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4441 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4501 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4561 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4621 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4681 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4741 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4801 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4861 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4921 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4981 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5041 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5101 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5161 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5221 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5281 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5341 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5401 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5461 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5521 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5581 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5641 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5701 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5761 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5821 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5881 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5941 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6001 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6061 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6121 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6181 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6241 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6301 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6361 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6421 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6481 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6541 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
