LOCUS       sequence-338379-        7439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of wildtype L. major glcyoprotein-63 from chromosome 31 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  AUTHORS   Antonia AL, Barnes AB, Martin AT, Wang L, Ko DC
  TITLE     Variation in Leishmania chemokine suppression driven by 
            diversification of the GP63 virulence factor.
  JOURNAL   PLoS Negl Trop Dis. 2021 Oct 28;15(10):e0009224. doi: 
            10.1371/journal.pntd.0009224. eCollection 2021 Oct.
  PUBMED    34710089
REFERENCE   2  (bases 1 to 7439)
  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: "PLoS Negl 
            Trop Dis."; date: "2021-10-28"; volume: "15(10):e0009224. doi"; 
            pages: " 10.1371/journal.pntd.0009224. eCollection 2021 Oct"
COMMENT     Sequence Label: Lmajor_gp63_31_2000_WT
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        149..528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        529..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     683..703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     777..796
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        777..795
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2631..2660
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2661..2678
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2775..2792)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2781..3005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3051..3479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3138..3157)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3348..3369
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3488..3508)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3493..3822
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3673..3808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3735..3754
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3871..4896
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3871..4896
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5026..5147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5063..5082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5117..5136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5196..5212)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5196..5212)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5209..5231)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5220..5236
                     /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(5244..5250,5251..5268,5269..5274))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5427..5444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5598..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5678..5697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6357..7148,7149..7217))
                     /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     6980..6999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7218..7322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7397..7416)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
       61 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
      121 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      721 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
      781 acgactcact atagggagac ccaagctggc tagcgtttaa acttaagctt ggtaccgcca
      841 ccatgtggtg gaggctgtgg tggttattac tgctgctgct gctgctgtgg ccaatggtgt
      901 gggcagcagc accaagctcc tgtggccact cccagatcgt gcacaagcag ggccacctgc
      961 ctctgacata cgccgccctg aggaacaatg gcaacggcct gcgctatcag gtggagatgg
     1021 acctggagag gtggaatggc agcgccaccg atgagcagag ggcagtggag ccacaggcaa
     1081 ccgtgacaat ctctagcgac cgggaggatt ctgagtgggc ccctatcaga atcgccgtgt
     1141 tcatgccaga cctggaggat gagagcaagt actgtacagc catcggccag atgagacccg
     1201 actcctctgg ctccctggtg ccttgcacct ctaacgccga cctgctgaca gatgccaaga
     1261 agaccgtgct gatcaacagc atcctgccac aggccgtgaa tatgcacgca cagcggctga
     1321 aggtgcggag acctgcagca tccacctcta tcgtggtgaa ttctatgcca ggcagctact
     1381 gtggctcctt tacagtgcca accgcacaca gaaccacagg cgtgcccgac gccgatttcg
     1441 tggtgtatgt ggcagcagga cccacctcta cacctaagag ctttatcgcc tgggccgtga
     1501 catgccagta ctatcctaac accgcaacag tgaccagcag gccagcagtg ggcgtgatgt
     1561 acttcaatcc aagatatctg cccacatccg agggcgagac ccaggagcac gtggacagat
     1621 atggcggcaa ccctagcggc tccaccaata ggctgaggag ggtggcatcc cacgagctgc
     1681 tgcacgccct gggcttcaca agctccgtgt ttaaggcaag gggaatgttt gcaaccgtgc
     1741 cctctctgag gggcaagagg aacgtgcctg tgctgaattc tagcgccgtg agagcagcag
     1801 caaaggcaaa gtacggcatc agcgacacag aggtgttcta tggagtggag ctggaggatc
     1861 agggagcacc aggcacctct ctgagccact ggaagcggag agccgccaag gatgagctga
     1921 tggcacccgt gttcggactg gcaaggtact cctctctgtc tctggccgcc ctggaggaca
     1981 tgggcttcta taaggtggac ttcagcaagg ccgagccagt ggccctggga gcaacagcag
     2041 gaggcaagct gttcaccgag ccatgcctga cagagggcac ctccaacaca cccaccgtgt
     2101 tttgcgactc cctgtctgcc agcgtgcgca gctgtacagc cgatcggctg tccatcggca
     2161 gatgcgccct gaccacatac agctccgccc tgcctagcta cgcccagtat ttccctaacc
     2221 agccaaccct gggcggctcc ctgtctcaca gcgactattg tcctgtgatc cagccactgt
     2281 ccaatacagg atgctccgga ggctctctgg gagcaatgcc aggctctatc accggcgata
     2341 acgccaggtg ttttgacgcc gataatctgc tggtgaagtt caccttcacc cagccaggag
     2401 caatctgcgc aaaggtgcac tgtaagaacg cctcccgcac ctacgaggtg ctggtcatcg
     2461 gcgccaatag ctggctgtcc tgcggagcag gaggcacagg agtgaccgtg cagccagccg
     2521 tgtctagccc taacgtgttc ctgaagggag gaaggatcgc atgcccacct tacgacgacg
     2581 tgtgctatgc caatccagtg gcctttgccg aggtggaggc accaccaacc gagcagaagc
     2641 tgatcagcga ggaggatctg caccaccacc accaccactg aggatccact agtccagtgt
     2701 ggtggaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta
     2761 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2821 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2881 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2941 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3001 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3061 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3121 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3181 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3241 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3301 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3361 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3421 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3481 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3841 atatccattt tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct
     3901 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     3961 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     4021 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     4081 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     4141 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     4201 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     4261 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     4321 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     4381 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     4441 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     4501 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     4561 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     4621 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     4681 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     4741 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     4801 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     4861 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tgctacgaga tttcgattcc
     4921 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4981 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5041 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5101 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5161 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5221 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5281 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5341 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5401 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6001 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7321 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7381 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctc
//
