LOCUS       sequence-227908-        7479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  AUTHORS   Wang ZY, Sato H, Kusam S, Sehra S, Toney LM, Dent AL
  TITLE     Regulation of IL-10 gene expression in Th2 cells by Jun proteins.
  JOURNAL   J Immunol. 2005 Feb 15;174(4):2098-105.
  PUBMED    15699140
REFERENCE   2  (bases 1 to 7479)
  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: "J 
            Immunol."; date: "2005-02-15"; volume: "174(4)"; pages: "2098-105"
COMMENT     Sequence Label: pMIEG3-c-Jun
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             145..661
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    725..1066
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1133..1549
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1479..1501
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1517..1533
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1575..1591
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1575..1591
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(1611..1622)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2609..2625)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2610..2626)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    join(2633..3207,3208..3210,3211..3219)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2800..2817)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3027..3046
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3220..3222,3223..3225,3226..3936)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3220..3222,3223..3225,3226..3936)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3265..3286)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3526..3545)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3873..3894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4018..4532
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4694..4710
                     /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(4718..4724,4725..4742,4743..4748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4763..4784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4901..4918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5072..5660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5152..5171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5831..6622,6623..6691))
                     /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     6454..6473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6692..6796)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6864..6882
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6920..6942)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7042..7061
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7255..7277
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7405..7424
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc tagttttgac
       61 tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat aaaagatttt
      121 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
      181 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
      241 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
      301 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
      361 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
      421 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
      481 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
      541 tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggact
      601 cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg ggtctttcat
      661 ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc ccccgccggg
      721 aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg
      781 catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac
      841 ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga
      901 ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc
      961 gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     1021 atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat
     1081 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1141 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1201 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1261 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1321 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1381 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1441 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1501 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg gaattccaag
     1561 gccttggcgg ccgctctaga actagtggat ccatgactgc aaagatggaa acgaccttct
     1621 atgacgatgc cctcaacgcc tcgttcctcc cgtccgagag cggaccttat ggctacagta
     1681 accccaagat cctgaaacag agcatgaccc tgaacctggc cgacccagtg gggagcctga
     1741 agccgcacct ccgcgccaag aactcggacc tcctcacctc gcccgacgtg gggctgctca
     1801 agctggcgtc gcccgagctg gagcgcctga taatccagtc cagcaacggg cacatcacca
     1861 ccacgccgac ccccacccag ttcctgtgcc ccaagaacgt gacagatgag caggagggct
     1921 tcgccgaggg cttcgtgcgc gccctggccg aactgcacag ccagaacacg ctgcccagcg
     1981 tcacgtcggc ggcgcagccg gtcaacgggg caggcatggt ggctcccgcg gtagcctcgg
     2041 tggcaggggg cagcggcagc ggcggcttca gcgccagcct gcacagcgag ccgccggtct
     2101 acgcaaacct cagcaacttc aacccaggcg cgctgagcag cggcggcggg gcgccctcct
     2161 acggcgcggc cggcctggcc tttcccgcgc aaccccagca gcagcagcag ccgccgcacc
     2221 acctgcccca gcagatgccc gtgcagcacc cgcggctgca ggccctgaag gaggagcctc
     2281 agacagtgcc cgagatgccc ggcgagacac cgcccctgtc ccccatcgac atggagtccc
     2341 aggagcggat caaggcggag aggaagcgca tgaggaaccg catcgctgcc tccaagtgcc
     2401 gaaaaaggaa gctggagaga atcgcccggc tggaggaaaa agtgaaaacc ttgaaagctc
     2461 agaactcgga gctggcgtcc acggccaaca tgctcaggga acaggtggca cagcttaaac
     2521 agaaagtcat gaaccacgtt aacagtgggt gccaactcat gctaacgcag cagttgcaaa
     2581 cattttgaga attcgatatc aagcttatcg ataccgtcga cctcgagatc cgcccctctc
     2641 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2701 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2761 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     2821 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     2881 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     2941 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     3001 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3061 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     3121 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3181 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     3241 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3301 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3361 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3421 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3481 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3541 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3601 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3661 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     3721 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     3781 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     3841 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     3901 gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc cgtcgacctg
     3961 cagccaagct tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     4021 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     4081 aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga
     4141 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4201 agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc
     4261 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4321 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4381 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa
     4441 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     4501 tgattgacta cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac
     4561 aacattctga gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg
     4621 aatccacata ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg
     4681 gagaattgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4741 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4801 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4861 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4921 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4981 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5041 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5101 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5161 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5221 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5281 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5341 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5401 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5461 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5521 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5581 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5641 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5701 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5761 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5821 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5881 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5941 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6001 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6061 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6121 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6181 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6241 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6301 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6361 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6421 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6481 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6541 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6601 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6661 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6721 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6781 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6841 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     6901 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6961 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     7021 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     7081 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     7141 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     7201 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     7261 cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     7321 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     7381 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     7441 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgta
//
