LOCUS       sequence-235023-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2473..2594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2510..2529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2564..2583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2601..3056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2738..2759)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2950..2969
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3083..3187
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3189..3546
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3189..3380
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3212..3232)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3397..3532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3581..4375
                     /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             3581..4375
                     /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(3635..3654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4245..4264
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4563..4582)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4607..4654
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4983..5571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5472..5491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttggt accgagctcg gatcgatatc tgcggccgcg
      661 tcgccaccat gtgtcctcag aagctaacca tctcctggtt tgccatcgtt ttgctggtgt
      721 ctccactcat ggccatgtgg gagctggaga aagacgttta tgttgtagag gtggactgga
      781 ctcccgatgc ccctggagaa acagtgaacc tcacctgtga cacgcctgaa gaagatgaca
      841 tcacctggac ctcagaccag agacatggag tcataggctc tggaaagacc ctgaccatca
      901 ctgtcaaaga gtttctagat gctggccagt acacctgcca caaaggaggc gagactctga
      961 gccactcaca tctgctgctc cacaagaagg aaaatggaat ttggtccact gaaattttaa
     1021 aaaatttcaa aaacaagact ttcctgaagt gtgaagcacc aaattactcc ggacggttca
     1081 cgtgctcatg gctggtgcaa agaaacatgg acttgaagtt caacatcaag agcagtagca
     1141 gttcccctga ctctcgggca gtgacatgtg gaatggcgtc tctgtctgca gagaaggtca
     1201 cactggacca aagggactat gagaagtatt cagtgtcctg ccaggaggat gtcacctgcc
     1261 caactgccga ggagaccctg cccattgaac tggcgttgga agcacggcag cagaataaat
     1321 atgagaacta cagcaccagc ttcttcatca gggacatcat caaaccagac ccgcccaaga
     1381 acttgcagat gaagcctttg aagaactcac aggtggaggt cagctgggag taccctgact
     1441 cctggagcac tccccattcc tacttctccc tcaagttctt tgttcgaatc cagcgcaaga
     1501 aagaaaagat gaaggagaca gaggaggggt gtaaccagaa aggtgcgttc ctcgtagaga
     1561 agacatctac cgaagtccaa tgcaaaggcg ggaatgtctg cgtgcaagct caggatcgct
     1621 attacaattc ctcatgcagc aagtgggcat gtgttccctg cagggtccga tccggcggcg
     1681 gcgggagtgg cggcgggggt tctggcggag gcctcgctag cggtggctcc atggtcagcg
     1741 ttccaacagc ctcaccctcg gcatccagca gctcctctca gtgccggtcc agcatgtgtc
     1801 aatcacgcta cctcctcttt ttggccaccc ttgccctcct aaaccacctc agtttggcca
     1861 gggtcattcc agtctctgga cctgccaggt gtcttagcca gtcccgaaac ctgctgaaga
     1921 ccacagatga catggtgaag acggccagag aaaaactgaa acattattcc tgcactgctg
     1981 aagacatcga tcatgaagac atcacacggg accaaaccag cacattgaag acctgtttac
     2041 cactggaact acacaagaac gagagttgcc tggctactag agagacttct tccacaacaa
     2101 gagggagctg cctgccccca cagaagacgt ctttgatgat gaccctgtgc cttggtagca
     2161 tctatgagga cttgaagatg taccagacag agttccaggc catcaacgca gcacttcaga
     2221 atcacaacca tcagcagatc attctagaca agggcatgct ggtggccatc gatgagctga
     2281 tgcagtctct gaatcataat ggcgagactc tgcgccagaa acctcctgtg ggagaagcag
     2341 acccttacag agtgaaaatg aagctctgca tcctgcttca cgccttcagc acccgcgtcg
     2401 tgaccatcaa cagggtgatg ggctatctga gctccgcctg agttaacgcg atcgcacgga
     2461 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2581 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2641 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2701 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2761 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2821 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2881 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2941 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3001 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3181 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3301 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3361 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3421 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3481 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3541 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3601 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3661 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3721 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3781 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3841 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3901 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3961 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4021 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4081 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4141 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4201 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4261 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4321 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4381 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4441 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4501 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4561 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4621 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4681 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4741 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4801 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4861 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4921 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4981 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5041 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5101 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5161 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5221 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5281 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5341 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5401 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5461 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5521 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5581 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5641 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
