LOCUS       sequence-204866-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of bioactive mouse IL-12 as a tandem dimer of p40 and p35
            linked with a Gly-Ser linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  AUTHORS   Nevil Singh
  TITLE     Nevil Singh IL-12 construct
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6955)
  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            "Unpublished"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: Tandem p40p35
FEATURES             Location/Qualifiers
     source          1..6955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          2802..3031
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3014..3033
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3089..3663,3664..3666,3667..3675)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3256..3273)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3483..3502
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3691..3700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3697..3699,3700..3702,3703..4416)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3742..3763)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4003..4022)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4350..4371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4442..4459)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4448..4672
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4719..4738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4798..4814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4798..4814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4811..4833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4822..4838
                     /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(4846..4852,4853..4870,4871..4876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4891..4912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5029..5046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5200..5788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5280..5299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5959..6750,6751..6819))
                     /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     6582..6601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6820..6924)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggccgcgtcg ccaccatgtg tcctcagaag ctaaccatct
      961 cctggtttgc catcgttttg ctggtgtctc cactcatggc catgtgggag ctggagaaag
     1021 acgtttatgt tgtagaggtg gactggactc ccgatgcccc tggagaaaca gtgaacctca
     1081 cctgtgacac gcctgaagaa gatgacatca cctggacctc agaccagaga catggagtca
     1141 taggctctgg aaagaccctg accatcactg tcaaagagtt tctagatgct ggccagtaca
     1201 cctgccacaa aggaggcgag actctgagcc actcacatct gctgctccac aagaaggaaa
     1261 atggaatttg gtccactgaa attttaaaaa atttcaaaaa caagactttc ctgaagtgtg
     1321 aagcaccaaa ttactccgga cggttcacgt gctcatggct ggtgcaaaga aacatggact
     1381 tgaagttcaa catcaagagc agtagcagtt cccctgactc tcgggcagtg acatgtggaa
     1441 tggcgtctct gtctgcagag aaggtcacac tggaccaaag ggactatgag aagtattcag
     1501 tgtcctgcca ggaggatgtc acctgcccaa ctgccgagga gaccctgccc attgaactgg
     1561 cgttggaagc acggcagcag aataaatatg agaactacag caccagcttc ttcatcaggg
     1621 acatcatcaa accagacccg cccaagaact tgcagatgaa gcctttgaag aactcacagg
     1681 tggaggtcag ctgggagtac cctgactcct ggagcactcc ccattcctac ttctccctca
     1741 agttctttgt tcgaatccag cgcaagaaag aaaagatgaa ggagacagag gaggggtgta
     1801 accagaaagg tgcgttcctc gtagagaaga catctaccga agtccaatgc aaaggcggga
     1861 atgtctgcgt gcaagctcag gatcgctatt acaattcctc atgcagcaag tgggcatgtg
     1921 ttccctgcag ggtccgatcc ggcggcggcg ggagtggcgg cgggggttct ggcggaggcc
     1981 tcgctagcgg tggctccatg gtcagcgttc caacagcctc accctcggca tccagcagct
     2041 cctctcagtg ccggtccagc atgtgtcaat cacgctacct cctctttttg gccacccttg
     2101 ccctcctaaa ccacctcagt ttggccaggg tcattccagt ctctggacct gccaggtgtc
     2161 ttagccagtc ccgaaacctg ctgaagacca cagatgacat ggtgaagacg gccagagaaa
     2221 aactgaaaca ttattcctgc actgctgaag acatcgatca tgaagacatc acacgggacc
     2281 aaaccagcac attgaagacc tgtttaccac tggaactaca caagaacgag agttgcctgg
     2341 ctactagaga gacttcttcc acaacaagag ggagctgcct gcccccacag aagacgtctt
     2401 tgatgatgac cctgtgcctt ggtagcatct atgaggactt gaagatgtac cagacagagt
     2461 tccaggccat caacgcagca cttcagaatc acaaccatca gcagatcatt ctagacaagg
     2521 gcatgctggt ggccatcgat gagctgatgc agtctctgaa tcataatggc gagactctgc
     2581 gccagaaacc tcctgtggga gaagcagacc cttacagagt gaaaatgaag ctctgcatcc
     2641 tgcttcacgc cttcagcacc cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct
     2701 ccgcctgagt taacgcgatc gcacggaatt cagtggatcc actagtaacg gccgccagtg
     2761 tgctggaatt aattcgctgt ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa
     2821 agcgggcatg acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt
     2881 cacctggccc gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag aaaagacaat
     2941 ctttttgttg tcaagcttga ggtgtggcag gcttgagatc tggccataca cttgagtgac
     3001 aatgacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca actgcaggtc
     3061 gagcatgcat ctagggcggc caattccgcc cctctccctc ccccccccct aacgttactg
     3121 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgtgattt tccaccatat
     3181 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     3241 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     3301 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     3361 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     3421 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     3481 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     3541 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     3601 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     3661 aagcttgcca caacccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
     3721 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3781 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3841 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
     3901 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3961 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4021 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4081 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     4141 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     4201 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     4261 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     4321 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4381 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccctagagc tcgctgatca
     4441 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4501 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4561 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4621 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4681 gcggaaagaa ccagctgggg ctcgagtgca ttctagttgt ggtttgtcca aactcatcaa
     4741 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5641 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
