LOCUS       sequence-262374-        7518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Leishmania infantum ectodomain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7518)
  AUTHORS   Roberts AJ, Ong HB, Clare S, Brandt C, Harcourt K, Franssen SU, 
            Cotton JA, Muller-Sienerth N, Wright GJ
  TITLE     Systematic identification of genes encoding cell surface and 
            secreted proteins that are essential for in vitro growth and 
            infection in Leishmania donovani.
  JOURNAL   PLoS Pathog. 2022 Feb 24;18(2):e1010364. doi: 
            10.1371/journal.ppat.1010364. eCollection 2022 Feb.
  PUBMED    35202447
REFERENCE   2  (bases 1 to 7518)
  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 
            Pathog."; date: "2022-02-24"; volume: "18(2):e1010364. doi"; pages: 
            " 10.1371/journal.ppat.1010364. eCollection 2022 Feb"
COMMENT     Sequence Label: D82_Li_04_0170
FEATURES             Location/Qualifiers
     source          1..7518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2682..2699
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2751..2806
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2805..2824)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5042..5060)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5128..5232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5233..5301,5302..6093)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5451..5470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6264..6852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6753..6772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7006..7023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7140..7161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7176..7181,7182..7199,7200..7206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7214..7230
                     /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)."
     primer_bind     7219..7241
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7238..7254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7238..7254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcct
     1201 acacaacgag tcaatgtcgg gaagtcagca gatgtgacgt agacggcacc ctggtgtacg
     1261 cgttctccgg ggagtgtgtc tgctgccccg agatgcccac aggagacatc ctgccagtac
     1321 aatgtgtccc atcaacagct cactgcccgt cagtacccag ctgtcgcgca tgtgatacaa
     1381 ccgcgggcat gtgtgacgcc tgtatgacag gctatgtgct ggaaccaaca aatatttgta
     1441 acgaatgtct ggcttccatg tgcgacgtct gccagtctag cagcgaaggt aagcgatgta
     1501 agctgtgctc cgttgggtac tattgggttt ctggcggtga gtgtcacgca gtgagcaact
     1561 cccaagctag cgagtctaat atgcagtgtc gcgtggcctt ttgcagcacc tgtgcgagca
     1621 ccactgcatc cacctgcgca aaatgcaacg acgggtacgc taaggactcc aacggatttt
     1681 gcgtttccga ctgtaatgtc caacagtgtg acatgtgtta taccgggaat agtgccgact
     1741 gtcagctctg ctccgccggc aacacctggt ctgccacggg atgttccgct ggagcctcag
     1801 gatgcagggt gggacactgc acaacttgcg tagcaggcga caacacaaat tgcgcgacat
     1861 gcgagtccgg atacacactc accgctgggt actgcctgcc acagcaactt tgtgcagatg
     1921 cgaattgcgc cagctgccca tccgacgcgg gtacttgtac acaatgcgcc aatggctatg
     1981 gtcttgtgga cggagcatgc gttcgatgtc aggttcctaa ctgtttctcc tgcgattctg
     2041 atgccaataa atgcgcccag tgcgcaccca actactatct gactccactg cttacttgca
     2101 gccccgtggc ttgtaatatc gaacactgca tgcaatgcga tcctcagact ccttcacggt
     2161 gccaagaatg tgtggcaccg tatgtggtgg attcctatga cggactgtgc aggctgagcg
     2221 atgcctgtag cgtccccaac tgtaagaaat gtgagacaga aacaagcaga ctgtgcgccg
     2281 agtgcgacac cggatacacc cttaccgccg acgcaacaag ctgtagctcc cctacgactc
     2341 agccctgcga cgtggagcac tgcaacacct gtgttaacgg agatagcacg aggtgcgctt
     2401 attgtaacac cgggtactac gtctctgatg gaaaatgcaa gaccatgcag gggtgttatg
     2461 tctccaattg tgcccagtgc atgctgctgg attcaactaa gtgctccaca tgcatgaaag
     2521 gttatctctt gacgagttcc tatggctgtg tgtcccagaa ggtcatcaac ggtgtagcag
     2581 cccctggcgc gcccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     2641 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     2701 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     2761 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2821 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2881 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2941 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     3001 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     3061 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     3121 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     3181 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     3241 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     3301 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3361 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3421 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3481 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3541 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3601 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3661 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3721 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3781 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3841 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3901 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3961 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     4021 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     4081 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     4141 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     4201 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     4261 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     4321 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4381 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4441 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4501 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4561 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4621 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4681 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4741 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4801 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4861 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4921 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4981 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     5041 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5101 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5161 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5221 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5281 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5341 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5401 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5461 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5521 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5581 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5641 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5701 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5761 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5821 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5881 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5941 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6001 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6061 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6121 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6181 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6241 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6301 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6361 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6421 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6481 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6541 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6601 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6661 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6721 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6781 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6841 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6901 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6961 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7021 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7081 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7141 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7201 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7261 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     7321 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     7381 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7441 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7501 catgtccaat atgaccgc
//
