LOCUS       sequence-337771-        6888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pET-15b(+) Toxoplasma gondii (delta prenylation motif).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  TITLE     Evolution of PRL
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6888)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pET-15b(+) Toxoplasma gondii (delta prenylatio...
FEATURES             Location/Qualifiers
     source          1..6888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1386..1404)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1400..1447
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1622..1627,1628..1644,1645..1650))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1677..1695)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1763..1867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1868..1936,1937..2728)
                     /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(2086..2105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2899..3487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3388..3407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3641..3658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3673..3815
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3741..3760)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3901..3923
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3917..4108)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5420..6502)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6464..6483
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6503..6580)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6786..6805
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg aacgaagcta gttcttgcct gtctccggaa
      181 gaaccgcgta gctgcgataa ccgtgaagaa ggtactaccc gtgaactgcg ttctggttcc
      241 cacatgggta aaggtaaact gggtcactct ccgaccacct ctcatagcga actgactcgt
      301 gaaggttcta gcgttgactg cggtcgtcgt ggtggtaacg aagaaaaaac tgaagaaaaa
      361 cgttctgaaa aagcaggtgc agaacgtgaa tgtgaacgta aaacccagcg tccgcgtttc
      421 gacggtgacg gttcttcgct gtccccgcgt agcaacgctt ctcgtggttc cgcttgctct
      481 acccgttcct gtgaatccca cgaaaaagta ggtcgtaccc tgtctggtga ttccaccggt
      541 cgtgacggcg tgtgctccgt atcttctgca cgtcagccga gcctgcgtcg taaatctgca
      601 gctgaagaag ttggtcgtga agttaaagtt gacggtaaag ctctggatag ccagatcgaa
      661 ctcgttcgta ccaaaaaccg tggtaaatct gaaggtgcac gcctgggtcg tggtgctggt
      721 tccgcgtgcg cggtttctgt ttctccgctg ggcaccgcgc gccacgtgcc gtctggtcgt
      781 cagccgtcgg aacgtcgtca gtctgcaaaa ccgctgacca aactgtcctc tagctcctct
      841 tctcacaaca aaactactat ggttccgact gttatgaaca ccccgacccg tatcgaagca
      901 ggtcgccaga aattcctgat cttcgacgcg ccgtcccagg aaaacctgcc ggcatacatt
      961 gaagaaatgc gcgcgtacga agtaaccgat ctggttcgta cctgtgaacg tacctacgac
     1021 gacaaaaccg ttctggcgtc tggcatccgt ccgcacgaac tgatttttcc ggacggcgaa
     1081 gcgccgccgg atgacgtgat cgatgaatgg ctgaccctgt gtaacgcggt ttctcagcag
     1141 cgtggtgcga tcgctatcca ctgcgtggcg ggtctgggcc gtgcgccggt gctggttgcg
     1201 atcgcactga tcgaaaaagg catggacccg atggacgcga tcatgttcat tcgtgaacgt
     1261 cgtaaaggcg ctattaaccg tcgtcagctg cagttcctga aaggctacaa acgtcgttct
     1321 aactataaaa aataaggatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1381 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1441 ttttttgctg aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc
     1501 ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca
     1561 ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg
     1621 cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct
     1681 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1741 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1801 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1861 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1921 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1981 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2041 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2101 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2161 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2221 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2281 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2341 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2401 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2461 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2521 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2581 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2641 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2701 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2761 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2821 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2881 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2941 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3001 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3061 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3121 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3181 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3241 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3301 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3361 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3421 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3481 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3541 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3601 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3661 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3721 catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3781 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3841 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3901 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3961 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4021 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4081 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4141 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4201 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4261 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4321 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4381 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4441 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4501 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4561 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     4621 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     4681 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     4741 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     4801 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     4861 ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt aggctggtaa gagccgcgag
     4921 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     4981 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     5041 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     5101 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     5161 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     5221 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     5281 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     5341 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     5401 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     5521 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     5581 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     5641 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     5701 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     5761 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     5821 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     5881 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     5941 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     6001 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     6061 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     6121 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     6181 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     6241 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     6301 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     6361 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     6421 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     6481 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     6541 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     6601 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     6661 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     6721 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     6781 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     6841 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
