LOCUS       sequence-197603-        5115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human IL-2 (C125S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5115)
  TITLE     Bacterial expression plasmid for production of histidine-tagged IL2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5115)
  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            "Unpublished"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: pQE80L-IL2 C125S
FEATURES             Location/Qualifiers
     source          1..5115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      427..521
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             565..1224
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(631..650)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1289..1375
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1468..2550)
                     /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     2512..2531
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2551..2628)
                     /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(2712..2734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2820..2960
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2875..2894
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2975..2992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3146..3734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3226..3245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3905..4696,4697..4765))
                     /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     4528..4547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4766..4870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4938..4956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(4980..4994,4995..5000,5001..5017,5018..5024)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    5000..5016
                     /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    5032..5048
                     /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             5067..5078
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             5097..5114
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gatccgcacc tacttcaagt tctacaaaga aaacacagct acaactggag catttactgc
       61 tggatttaca gatgattttg aatggaatta ataattacaa gaatcccaaa ctcaccagga
      121 tgctcacatt taagttttac atgcccaaga aggccacaga actgaaacat cttcagtgtc
      181 tagaagaaga actcaaacct ctggaggaag tgctaaattt agctcaaagc aaaaactttc
      241 acttaagacc cagggactta atcagcaata tcaacgtaat agttctggaa ctaaagggat
      301 ctgaaacaac attcatgtgt gaatatgctg atgagacagc aaccattgta gaatttctga
      361 acagatggat taccttttcc caaagcatca tctcaacact gacttaagct taattagctg
      421 agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
      481 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag cttggcgaga
      541 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
      601 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
      661 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
      721 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
      781 tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
      841 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
      901 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
      961 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     1021 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     1081 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     1141 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     1201 gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct
     1261 ggggtaatga ctctctagct tgaggcatca aataaaacga aaggctcagt cgaaagactg
     1321 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1381 ctctagatta cgtgcagtcg atgataagct gtcaaacatg agaattgtgc ctaatgagtg
     1441 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1501 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1561 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     1621 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     1681 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     1741 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     1801 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     1861 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     1921 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     1981 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     2041 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     2101 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     2161 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     2221 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     2281 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     2341 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     2401 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     2461 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     2521 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     2581 gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcgg aatttcgggc
     2641 agcgttgggt cctggccacg ggtgcgcatg atctagagct gcctcgcgcg tttcggtgat
     2701 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2761 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2821 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     2881 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2941 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4921 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagaaat
     4981 cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca attgtgagcg
     5041 gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga ggatcgcatc
     5101 accatcacca tcacg
//
