LOCUS       sequence-369718-        5058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human IL-15 with N-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5058)
  TITLE     McLellan Lab Plasmids 2022
REFERENCE   2  (bases 1 to 5058)
  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"
COMMENT     Sequence Label: pQE80L Hu IL-15-His tag
FEATURES             Location/Qualifiers
     source          1..5058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      370..464
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             508..1167
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(574..593)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1232..1318
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1411..2493)
                     /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     2455..2474
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2494..2571)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(2655..2677)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2763..2903
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2818..2837
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2918..2935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3089..3677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3848..4639,4640..4708))
                     /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     4471..4490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4709..4813)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4881..4899
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4923..4937,4938..4943,4944..4960,4961..4967)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    4943..4959
                     /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)."
     protein_bind    4975..4991
                     /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             5010..5021
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5040..5057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gatccaattg ggtgaacgtg atttccgact tgaaaaaaat cgaggacctg atccaatcta
       61 tgcacataga cgctaccctc tacacggaat cagatgtgca tccatcctgt aaagtgacag
      121 cgatgaagtg cttcctcttg gaacttcaag tcataagcct ggaatctggt gatgcaagta
      181 tacatgatac tgtcgaaaat ctgattatac tcgccaataa ctctttgagt agtaatggta
      241 atgttaccga gtctggctgt aaagaatgtg aggaactcga ggaaaagaat attaaagaat
      301 ttttgcagag cttcgtgcat atagttcaga tgttcatcaa taccagctaa gcttaattag
      361 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
      421 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg
      481 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
      541 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
      601 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
      661 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
      721 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
      781 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
      841 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
      901 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
      961 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1021 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1081 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1141 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     1201 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1261 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1321 gccctctaga ttacgtgcag tcgatgataa gctgtcaaac atgagaattg tgcctaatga
     1381 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1441 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1501 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     1561 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     1621 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     1681 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     1741 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     1801 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     1861 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     1921 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     1981 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     2041 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     2101 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     2161 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     2221 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     2281 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     2341 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     2401 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     2461 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     2521 cgggcgctat catgccatac cgcgaaaggt tttgcaccat tcgatggtgt cggaatttcg
     2581 ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga gctgcctcgc gcgtttcggt
     2641 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2701 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2761 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     2821 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2881 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4801 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4861 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgaga
     4921 aatcataaaa aatttatttg ctttgtgagc ggataacaat tataatagat tcaattgtga
     4981 gcggataaca atttcacaca gaattcatta aagaggagaa attaactatg agaggatcgc
     5041 atcaccatca ccatcacg
//
