LOCUS       sequence-310420-        7847 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses N-terminally HA-tagged PfROM4 variant G470A/G471N (codon 
            optimized for mammalian expression) from pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7847)
  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 7847)
  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 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 (G470A/G471N)
FEATURES             Location/Qualifiers
     source          1..7847
                     /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             967..993
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1003..1029
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1039..1065
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3438..3455)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3444..3668
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3714..4142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3801..3820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4011..4032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4151..4171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4156..4485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4336..4471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4398..4417
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4552..5346
                     /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(4606..4625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5216..5235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5520..5641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5557..5576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5611..5630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5690..5706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5690..5706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5703..5725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5714..5730
                     /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(5738..5744,5745..5762,5763..5768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5783..5804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5921..5938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6092..6680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6172..6191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6851..7642,7643..7711))
                     /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     7474..7493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7712..7816)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcaag
      961 atggactacc cttacgatgt cccagactac gctggcggag gatatccata tgatgtgcct
     1021 gattacgctg gcgggggata tccttatgac gtgccagatt atgccggagg agggggcagc
     1081 aataccggct tcaaccccaa gaacgccgag aagaagcgga gcagcatctt cagcgacctg
     1141 gagatccccc agggcgacgt gaagaagaag gcctccatct tcaataatat ggagaagaaa
     1201 aagatgacca tcttcaagaa aaccaagggc gagaacaacg aggagaagaa gaaaagaagc
     1261 atcttcaata acaataataa caacaataat aataacaaca ccatgaagat caacatcttt
     1321 aacaataaca acgagaagaa agaaagcttc tccattaacg gggatgaaaa gaagaaatcc
     1381 ttttccataa atggcgacga gaaaaaggag tctttttcca ttaacggaga tgaaaaaaag
     1441 gagagcttcc tgatcaacaa agacgagaaa aaagagtctt tcctgattaa taaggatgaa
     1501 aaggagaaga gcttctctat caatggcgac gaaaagaaag agagcttttc tattaacggg
     1561 gatgaaaaga aggaatcctt tagcatcaat gaggacgaga ccaagaagag ctgttccatc
     1621 aacgacgatg agcggaagat ctccatcttc tccaataacg aaaagaagaa gaacagcatc
     1681 tactccgaca cccagagcag caagcgggag gatgagaagc ggatctccat ttttagcgat
     1741 ctggagacca gcaccaacat cgacgacatc agctccaagc ggaacagcca gaagctgtcc
     1801 gtgtacggcg acaacaagct gaagaagggc agcctgctgt cccctaaggt ggacaactac
     1861 cggaacacca tcgacaacat caacgagatc aacgacatca agatcatcgt gaccagcgac
     1921 gagaatctgc acaccctgcc cagcggagct gtgggcagaa gagcccccct gaaccctttc
     1981 agcagcccca tcctgggcaa gtaccggcgg aagaacaaga acgccaaggc caaggtgaag
     2041 gacccccggc tgaacaacaa tcccctgatc ggcaggctga ccgtgtgtat cagcaccacc
     2101 gccatcctgt tctgggtgtt cttcgccgag atggtgttca actacaacac cttcaacggc
     2161 cggtgtatct ccaaggtgct gtaccccatc tacaccgaga acgtggtgct gaaacggcag
     2221 cccttcttcg tgttcctggg ctacggcgcc tgtgagtaca acctggacga gagcgccagc
     2281 aacagacact tcatcggcag caaggccagc gatgagggct ggcctggcga caaggtggag
     2341 gagaaccctg atggcagagg ctacgccaac tgggacagcg tgaacaaccg ggtgtacaat
     2401 ctgctgggcg gcctgaacac caactacatc cggaactacg gcgagctgta caggctgttc
     2461 tggagcatgt acctgcacgc caacttcatg cacatcctgt tcaacgtgat ctgccagatc
     2521 cagatcctgt ggatgatcga gcctgactgg ggcagcatca gaaccggcct gctgttcttt
     2581 attagcggcg tgaccggcaa tctgctgtcc gccgtgtgtg acccctgtgg cgtgaccatc
     2641 ggcagctccg gcagcctgta cggcctgatc ggcgccctgt tcgcctacta catcgagtac
     2701 tggaaaacca tccccaggcc ctgctgtgtg ctgatcttca tgtttctggt ggtgatgttc
     2761 ggcatcattg tgggcatgtt cggctacacc gacaactacg cccacatcgg cggctgcctg
     2821 ggcggcgtgc tgttcggatt cagcacaatc accaccgtgt ctgccgccga taagtgtacc
     2881 ctgggcgaga gaatgctggt gtccgccccc ttcagctggt tcctgagcaa tgagaccaag
     2941 gagctgatca tcgccaaagc caaggacaag aagatcaagg gggagaactt ccggaagaag
     3001 cagctggcca acaaggtgca caagaacgac gccctgcacg tggccatggc tgtcatgaag
     3061 aaccggatca acgatgaggg cagacctcct tgccggatga agctgaggga gtggattgtg
     3121 agaatcaccg ccgccagcac cctgatcatc atgtggatcg tgctgttcat ctacctgctg
     3181 aacgagaagg cctacaagag ctacagcccc ctgggccaga tcaagtttag cggcgtgcac
     3241 agctgctact gctgccagat cgtgaagaac aagttcacct acatcaaggt gaacgacttc
     3301 tactggtgct tcaccaccga ggaggccacc cggtactact gtaacaagtg atagtctaga
     3361 acgcggagct ccagcttttg ttccctttag tgagcggccg ctcgagtcta gagggcccgt
     3421 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3481 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3541 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3601 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3661 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3721 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3781 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3841 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3901 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3961 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4021 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4081 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4141 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4201 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4321 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4381 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4441 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4501 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4561 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4621 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4681 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4741 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4801 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4861 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4921 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4981 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5041 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5101 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5161 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5221 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5281 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5341 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5401 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5461 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5521 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5581 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5641 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5701 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5761 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5821 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5881 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5941 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6001 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6061 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6121 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6421 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6481 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6541 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6601 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6721 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6781 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6841 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6901 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6961 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7021 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7081 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7141 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7201 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7261 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7321 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7381 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7441 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7501 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7561 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7621 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7681 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7741 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
