LOCUS       sequence-338383-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of wildtype L. major glcyoprotein-63 from chromosome 28 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  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 7259)
  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_28_0570_WT
FEATURES             Location/Qualifiers
     source          1..7259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..531
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        532..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     780..799
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        780..798
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1815..1824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2454..2483
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2484..2501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2598..2615)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2604..2828
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2874..3302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2961..2980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3171..3192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3311..3331)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3316..3645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3496..3631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3558..3577
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3694..4719
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3694..4719
                     /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    4849..4970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4886..4905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4940..4959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5019..5035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5019..5035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5032..5054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5043..5059
                     /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(5067..5073,5074..5091,5092..5097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5112..5133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5250..5267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5421..6009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5501..5520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6180..6971,6972..7040))
                     /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     6803..6822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7041..7145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7220..7239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
       61 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      121 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
      181 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      241 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      301 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      361 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      421 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      481 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      541 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      601 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      661 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
      781 aatacgactc actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg
      841 ccaccatgtg gtggaggctg tggtggttat tactgctgct gctgctgctg tggcctatgg
      901 tgtgggcaca gggacaccca gagagggcag actccgagga gccaagatgc ggctttgatg
      961 agctggaggc ccacaccatc ggcacaaggg tgtctggaat cagcagggtg gagctgccaa
     1021 ccggagagct ggtggtggca gcagcagcaa caggcgccct gcagccaatc aggatcgccg
     1081 tgttcaccga cgatatctcc aacagctccc agcactgcac agcatccgga cagtctaggc
     1141 caaacttccg gggctccagg gtgacctgtt ctgccgcaga ggtgctgaca cgggccaaga
     1201 agagagtgct gctggagctg ctgatcccaa gcgccgtgca gctgcaccag gagaggctga
     1261 acgtgcagcg cgtgaacggc aatatcgtgg tggactctag catccagaag gatcgggtgt
     1321 gcggccagtt ctctatcaga gaggagcaca tgaagaccgg cgtgaaggac gccgattttg
     1381 tgctgtacat gagcgccgcc cctacatctg gcagcgtgat cgcctgggcc ctgaagtgtc
     1441 agaacttcga caatggccgg ccatccgtgg gcgtggtgaa catctctcca aagtatatcg
     1501 ccgccgatcc caagaccgtg agagtgatcg cacacgaggt gctgcacgca ctgggcttct
     1561 cccggagcgt gttccaggag agaaatatgc tggcaatggc aagcttccgg tccaagggac
     1621 caagccccgt gatctgctcc gagaaggtgg tggccaaggc ccagcagcac tacggctgta
     1681 agacccaggc ctttatggag ctggaggaca caggcgatat cgacgatgcc tcctctcact
     1741 ggaagcggag aaatgccaag gacgagctga tggcaggctt ctccggagtg ggcatctatt
     1801 ctgccctgac catcgccgcc atggaggata caggctacta tcagggcaac tacgccaagg
     1861 cagagccaat ggcatatgga cacgacgcag gatgcaagct gagctccgat cagtgcgtga
     1921 ccaatagcac atcccagatc cctggcatgt tttgcgacgc ccccgatgcc ccttggtctt
     1981 gtaccagcga caggctggga gtgggcagat gcatcctgac ctcccacaag tctaacctgc
     2041 ctacatactt ccagtatttt tccgatccaa gactgggcgg ccctgaccca ctgatggatt
     2101 tttgccctgt ggtggaggtg gcagagggca ccatgtgcgc agccaccaca aatgccctga
     2161 agggcagcgt gtacggcgtg atgtccagat gcgtggacac acccgtgggc ttctctatgg
     2221 acgatagcgc cgtgcgccag cacggaatct gcgtggaggt gcagtgtgac agcaccaagt
     2281 actatatcaa ggccaacgga gcatccgcct tctgcgattg tccacctggc tctacctaca
     2341 atctgagcac actgtctccc agcttttcca agggctacct ggtgtgccct agctatgaga
     2401 gcgtgtgcgc catcaagatc aacgcctctc tgtacgagga gtatagcggc ttcgagcaga
     2461 agctgatcag cgaggaggac ctgcaccacc accaccacca ctgaggatcc actagtccag
     2521 tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt
     2581 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2641 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2701 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2761 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2821 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2941 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3001 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3061 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3181 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3241 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3301 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3361 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3481 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3541 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3601 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3661 tgtatatcca ttttcggatc tgatcagcac gtgatgaaaa agcctgaact caccgcgacg
     3721 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3781 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3841 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3901 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     3961 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     4021 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     4081 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     4141 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4201 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     4261 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4321 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4381 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4441 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     4501 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4561 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4621 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4681 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtgctacg agatttcgat
     4741 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4801 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4861 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4921 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4981 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5041 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5101 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5161 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5221 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5281 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5341 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5401 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5461 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5521 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5581 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5641 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5701 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5761 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5821 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5881 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5941 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6001 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6061 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6121 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6181 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6241 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6301 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6361 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6421 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6481 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6541 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6601 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6661 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6721 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6781 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6841 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6901 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6961 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7021 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7081 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7141 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7201 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctg
//
