LOCUS       sequence-46609-e        7295 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 7295)
  AUTHORS   Li S, Wang L, Berman M, Kong YY, Dorf ME
  TITLE     Mapping a dynamic innate immunity protein interaction network 
            regulating type I interferon production.
  JOURNAL   Immunity. 2011 Sep 23;35(3):426-40. Epub 2011 Sep 8.
  PUBMED    21903422
REFERENCE   2  (bases 1 to 7295)
  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: "Immunity.";
            date: "2011-09-23"; volume: "35(3)"; pages: "426-40. Epub 2011 Sep 
            8"
COMMENT     Sequence Label: mMib1-FLAG
FEATURES             Location/Qualifiers
     source          1..7295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3720..3743
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3796..3815)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3797..3815)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4089..4210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4126..4145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4180..4199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4217..4672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4354..4375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4566..4585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4699..4803
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4805..5162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4805..4996
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4828..4848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5013..5148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5197..5991
                     /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             5197..5991
                     /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(5251..5270)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5861..5880
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6179..6198)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6223..6270
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6599..7187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7088..7107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgctctagc ccgggcggat ccaccatgag taactcccgc aacaaccgag
      721 tgatggtgga gggggtcggc gcccgcgtag tgcgcggccc ggactggaag tggggcaagc
      781 aggacggcgg cgagggccat gtaggcactg tacggagctt cgagagtccg gaggaggtcg
      841 tggtggtgtg ggacaacggc acggccgcca actaccgctg ctcaggggcc tacgacctgc
      901 gtatcctgga cagcgcgccc accggcatca agcatgatgg aactatgtgt gatacctgcc
      961 gccagcaacc aatcattggc attcgatgga aatgtgcaga gtgcaccaat tatgatttgt
     1021 gcactgtttg ttatcatgga gataaacatc acttaaggca tcgcttttat aggattacta
     1081 caccaggaag tgaacgggtt ctgttagagt ctcgtagaaa atctaagaag attacagcca
     1141 gaggaatctt tgctggtgcc agagtggtgc gaggagtgga ctggcagtgg gaagatcaag
     1201 atggaggcaa tgggcgtaga ggaaaggtca cggagatcca ggactggagt gcatccagcc
     1261 cacacagtgc agcctatgtt ctctgggata acggtgctaa gaacctttac agggttggct
     1321 ttgagggcat gtctgatctc aagtgtgtcc aggatgccaa aggaggttct ttctacagag
     1381 atcattgccc tgtgctaggt gagcagaatg gcaacaggaa tcctggtgga ttgcagattg
     1441 gtgacctggt aaatatagac cttgatctag aaattgtcca gtctttacag cacggtcatg
     1501 gaggatggac cgatggcatg tttgagactt taactacaac tggaaccgtc tgtggcattg
     1561 atgaagatca tgacattgta gtacagtatc caagtggcaa tagatggact ttcaatcctg
     1621 ctgttctcac taaggcaaac attgtccgaa gtggggatgc tgcacagggt gcagaaggag
     1681 gcacctcaca gtttcaagtg ggtgatcttg tgcaagtttg ttatgatctg gaaagaatta
     1741 aacttctcca aagaggacat ggtgaatggg ctgaagcaat gcttccaact ttaggtaaag
     1801 ttggccgagt acaacagatt tattcggaca gtgacttaaa ggtggaagtc tgtggaacat
     1861 cttggacgta taaccctgca gcagtttcca aggtggcccc tgcaggatca gccattagca
     1921 atgcatctgg tgaaagactc tcccagcttc tgaagaaatt atttgaaacc caagaatctg
     1981 gtgacctcaa tgaagagctg gttaaggctg ctgccaatgg agacgtggct aaagtggaag
     2041 atttgctgaa aagaccagat gtggatgtaa acggacagtg tgctggccac acagcaatgc
     2101 aagctgctag tcagaacgga catgttgata ttttgaaatt acttttgaag caaaatgttg
     2161 atgttgaagc agaggataaa gatggagatc gagccgttca ccatgcagcc tttggagatg
     2221 agggtgctgt catagaagtc ctacaccgag gcagtgctga cctgaatgct cgtaataagc
     2281 gtagacagac gcctcttcat attgctgtca ataaaggtca tcttcaagtt gtgaagactt
     2341 tgttggattt tggttgtcat cccagtctcc aggattctga aggtgacacc cctctccatg
     2401 atgccataag taagaaacgg gatgacattc tggcagttct tctggaagct ggagcagatg
     2461 tgaccatcac gaacaataac gggttcaatg ccctgcacca tgctgcactg agagggaacc
     2521 ccagtgcaat gcgtgtttta ctctcaaaat taccaagacc atggattgtg gatgagaaga
     2581 aagatgatgg ttatactgcc ttgcacctgg ctgccctgaa taatcatgtg gaagtggctg
     2641 agctgttggt gcatcaggga aatgcaaacc tggatatcca gaatgtgaac cagcaaacag
     2701 ccctacacct tgctgttgaa cggcaacaca cccagattgt taggcttttg gtccgtgcag
     2761 gtgctaagct agacattcag gataaggatg gagatactcc tctgcatgaa gctttgagac
     2821 atcatacttt gtctcaacta cgtcagctac aagacatgca agatgtgggc aaggtagacg
     2881 ctgcctggga accttcaaaa aacacattaa taatgggact tggtacccag ggggctgaga
     2941 agaagagtgc agcatccatt gcgtgtttct tggcagccaa tggtgctgat ctcagcattc
     3001 gaaataagaa gggtcagtca ccactggatc tctgtcctga cccaagtctc tgcaaagctt
     3061 tggcaaaatg tcataaggaa aaagtcagtg gccaagtggg ttctcggagt ccttctatga
     3121 ttagcaacga ttctgaaact ttagaggaat gtatggtgtg ctcagacatg aagagagata
     3181 ctctttttgg tccgtgtgga cacattgcaa cctgttcttt gtgttcccct cgtgtcaaga
     3241 aatgcctcat ttgtaaagag caggttcagt ccaggacaaa gattgaagaa tgtgtggtgt
     3301 gctccgacaa gaaagcagct gttctctttc aaccttgtgg gcacatgtgt gcttgtgaga
     3361 actgtgccag cctgatgaaa aagtgtgtgc agtgtcgggc agtggttgag cggagggtgc
     3421 ctttcatcac gtgctgtggt ggcaagagct cagaagaccc ctctgatgaa atctcaagtg
     3481 ggaatatccc agtgctgcag aaggacaaag acaacaccaa tgttaacgca gatgtgcaga
     3541 aactgcagca gcagcttcag gacattaagg agcagaccat gtgccctgtg tgcttggatc
     3601 gcctgaagaa catgattttc ctctgtggcc atggaacatg ccagctctgt ggagaccgga
     3661 tgagtgaatg cccaatttgt cgcaaggcaa ttgaacgaag gattcttttg tacctcgagg
     3721 attacaagga tgacgacgat aagtagggcc cggtacctta attaattaag gtaccaggta
     3781 agtgtaccca attcgcccta tagtgagtcg tattacaatt cactcgatcg cccttcccaa
     3841 cagttgcgca gcctgaatgg cgaatggaga tccaattttt aagtgtataa tgtgttaaac
     3901 tactgattct aattgtttgt gtattttaga ttcacagtcc caaggctcat ttcaggcccc
     3961 tcagtcctca cagtctgttc atgatcataa tcagccatac cacatttgta gaggttttac
     4021 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     4081 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4141 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4201 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4261 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4321 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4381 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4441 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4501 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4561 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4621 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4681 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4741 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4801 aatcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4861 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4981 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5041 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5101 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     5161 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5221 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5281 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5341 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     5401 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5461 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5521 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5581 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5641 gaagccggtc ttgtcgatca ggatgatctg gacgaagaac atcaggggct cgcgccagcc
     5701 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5761 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5821 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5881 gctgaagaac ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5941 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6001 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6061 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6121 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     6181 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     6241 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     6301 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     6361 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     6421 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     6481 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6541 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6601 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6661 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6721 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6781 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6841 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6901 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6961 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7021 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7081 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7141 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7201 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7261 tatcccctga ttctgtggat aaccgtatta ccgcc
//
