LOCUS       sequence-238577-        5685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of murine cytokine IL-12 under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5685)
  AUTHORS   Tzeng SY, Patel KK, Wilson DR, Meyer RA, Rhodes KR, Green JJ
  TITLE     In situ genetic engineering of tumors for long-lasting and systemic 
            immunotherapy.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Feb 7. pii: 1916039117. doi: 
            10.1073/pnas.1916039117.
  PUBMED    32034097
REFERENCE   2  (bases 1 to 5685)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Feb 7. pii: 1916039117. doi: 
            10.1073/pnas.1916039117."
COMMENT     Sequence Label: mIL12-N1
FEATURES             Location/Qualifiers
     source          1..5685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2413..2534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2450..2469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2504..2523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2541..2996)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2678..2699)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2890..2909
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3023..3127
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3129..3486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3129..3320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3152..3172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3337..3472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3521..4315
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3521..4315
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3575..3594)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4185..4204
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4503..4522)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4547..4594
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4923..5511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5412..5431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttggt accgagctcg gatcgatatc tgcggccgcg
      601 tcgccaccat gtgtcctcag aagctaacca tctcctggtt tgccatcgtt ttgctggtgt
      661 ctccactcat ggccatgtgg gagctggaga aagacgttta tgttgtagag gtggactgga
      721 ctcccgatgc ccctggagaa acagtgaacc tcacctgtga cacgcctgaa gaagatgaca
      781 tcacctggac ctcagaccag agacatggag tcataggctc tggaaagacc ctgaccatca
      841 ctgtcaaaga gtttctagat gctggccagt acacctgcca caaaggaggc gagactctga
      901 gccactcaca tctgctgctc cacaagaagg aaaatggaat ttggtccact gaaattttaa
      961 aaaatttcaa aaacaagact ttcctgaagt gtgaagcacc aaattactcc ggacggttca
     1021 cgtgctcatg gctggtgcaa agaaacatgg acttgaagtt caacatcaag agcagtagca
     1081 gttcccctga ctctcgggca gtgacatgtg gaatggcgtc tctgtctgca gagaaggtca
     1141 cactggacca aagggactat gagaagtatt cagtgtcctg ccaggaggat gtcacctgcc
     1201 caactgccga ggagaccctg cccattgaac tggcgttgga agcacggcag cagaataaat
     1261 atgagaacta cagcaccagc ttcttcatca gggacatcat caaaccagac ccgcccaaga
     1321 acttgcagat gaagcctttg aagaactcac aggtggaggt cagctgggag taccctgact
     1381 cctggagcac tccccattcc tacttctccc tcaagttctt tgttcgaatc cagcgcaaga
     1441 aagaaaagat gaaggagaca gaggaggggt gtaaccagaa aggtgcgttc ctcgtagaga
     1501 agacatctac cgaagtccaa tgcaaaggcg ggaatgtctg cgtgcaagct caggatcgct
     1561 attacaattc ctcatgcagc aagtgggcat gtgttccctg cagggtccga tccggcggcg
     1621 gcgggagtgg cggcgggggt tctggcggag gcctcgctag cggtggctcc atggtcagcg
     1681 ttccaacagc ctcaccctcg gcatccagca gctcctctca gtgccggtcc agcatgtgtc
     1741 aatcacgcta cctcctcttt ttggccaccc ttgccctcct aaaccacctc agtttggcca
     1801 gggtcattcc agtctctgga cctgccaggt gtcttagcca gtcccgaaac ctgctgaaga
     1861 ccacagatga catggtgaag acggccagag aaaaactgaa acattattcc tgcactgctg
     1921 aagacatcga tcatgaagac atcacacggg accaaaccag cacattgaag acctgtttac
     1981 cactggaact acacaagaac gagagttgcc tggctactag agagacttct tccacaacaa
     2041 gagggagctg cctgccccca cagaagacgt ctttgatgat gaccctgtgc cttggtagca
     2101 tctatgagga cttgaagatg taccagacag agttccaggc catcaacgca gcacttcaga
     2161 atcacaacca tcagcagatc attctagaca agggcatgct ggtggccatc gatgagctga
     2221 tgcagtctct gaatcataat ggcgagactc tgcgccagaa acctcctgtg ggagaagcag
     2281 acccttacag agtgaaaatg aagctctgca tcctgcttca cgccttcagc acccgcgtcg
     2341 tgaccatcaa cagggtgatg ggctatctga gctccgcctg agttaacgcg atcgcacgga
     2401 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2521 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2581 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2641 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2701 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2761 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2821 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2881 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2941 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3001 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3061 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3121 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3421 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3481 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3541 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3601 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3661 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3721 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3781 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3841 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3901 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3961 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4021 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4081 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4141 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4201 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4261 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4321 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4381 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4441 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4501 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4561 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4621 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4681 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4741 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4801 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4861 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5581 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     5641 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
