LOCUS       sequence-12947-e        7062 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 7062)
  AUTHORS   Miyamichi K, Amat F, Moussavi F, Wang C, Wickersham I, Wall NR, 
            Taniguchi H, Tasic B, Huang ZJ, He Z, Callaway EM, Horowitz MA, Luo 
            L
  TITLE     Cortical representations of olfactory input by trans-synaptic 
            tracing.
  JOURNAL   Nature. 2011 Apr 14;472(7342):191-6. doi: 10.1038/nature09714.
  PUBMED    21179085
REFERENCE   2  (bases 1 to 7062)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2011-04-14- 14"; volume: "472"; issue: "7342"; 
            pages: "191-6"
COMMENT     Sequence Label: pAAV-TRE-HTG
FEATURES             Location/Qualifiers
     source          1..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        159..473
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    203..221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    238..256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    274..292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    310..328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    381..399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     446..470
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             495..872
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      885..894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(891..893,894..896,897..1607)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(891..893,894..896,897..1607)
                     /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(936..957)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1197..1216)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1544..1565
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1617..1682
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             2163..2216
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    3809..4285
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4008..4027)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4325..4465
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4325..4454
                     /label=AAV2 ITR
     rep_origin      4540..4995
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4627..4646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4837..4858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5012..5031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5131..5153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5191..5209)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5277..5381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5382..5450,5451..6242)
                     /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     complement(5600..5619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6413..7001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6902..6921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctcga gtttactccc tatcagtgat
      181 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc
      241 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      301 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag
      361 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
      421 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
      481 ttctagccgc caccatgcca gagccagcga agtctgctcc cgccccgaaa aagggctcca
      541 agaaggcggt gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg
      601 agagctattc catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt
      661 cgtccaaggc catgggcatc atgaattcgt ttgtgaacga cattttcgag cgcatcgcag
      721 gtgaggcttc ccgcctggcg cattacaaca agcgctcgac catcacctcc agggagatcc
      781 agacggccgt gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta
      841 ctaaggccat caccaagtac accagcgcta aggatccacc ggtcgccacc atggtgagca
      901 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      961 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1021 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1081 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1141 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1201 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1261 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1321 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1381 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1441 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1501 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1561 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt agtggtgtga
     1621 aacagacttt gaattttgac cttctcaagt tggcgggaga cgtggagtcc aacccagggc
     1681 ccatggcgcg gctgctgccc gcgctgctgc tgctgctgct gcccggtaac gtgaccggta
     1741 acgggtccgg taacggttct ttgtcccgtt gcccccccgg tcagttccgc tgctcggagc
     1801 cgcccggtgc ccacggggag tgttacccgc aggactggct gtgcgacgga caccccgact
     1861 gcgacgacgg gcgggacgag tggggctgcg ggaccagcgc gatccccgcg gtgcccacgg
     1921 acaacggcac agaggctccc actgcccctg ctcctggacg tgctctgcca gccaggaatc
     1981 acggccgcat gtggatgctg atcactgcag tgctcctgtg ctgcctggta gctgtgggtg
     2041 gtatcgctgc atgggggaag tccaaagcaa aaagcaggtc tgacatcttc agtcttgaaa
     2101 gcgcatccaa ggagctgctg gtgcctgaca agagccaggc agacttgttc tccggtagtg
     2161 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg
     2221 ttcctcaggc tctcctgttt gtaccccttc tggtttttcc attgtgtttt gggaaattcc
     2281 ctatttacac gataccagac aagcttggtc cctggagtcc gattgacata catcacctca
     2341 gctgcccaaa caatttggta gtggaggacg aaggatgcac caacctgtca gggttctcct
     2401 acatggaact taaagttgga tacatcttag ccataaaagt gaacgggttc acttgcacag
     2461 gcgttgtgac ggaggctgaa acctacacta acttcgttgg ttatgtcaca accacgttca
     2521 aaagaaagca tttccgccca acaccagatg catgtagagc cgcgtacaac tggaagatgg
     2581 ccggtgaccc cagatatgaa gagtctctac acaatccgta ccctgactac cgctggcttc
     2641 gaactgtaaa aaccaccaag gagtctctcg ttatcatatc tccaagtgtg gcagatttgg
     2701 acccatatga cagatccctt cactcgaggg tcttccctag cgggaagtgc tcaggagtag
     2761 cggtgtcttc tacctactgc tccactaacc acgattacac catttggatg cccgagaatc
     2821 cgagactagg gatgtcttgt gacattttta ccaatagtag agggaagaga gcatccaaag
     2881 ggagtgagac ttgcggcttt gtagatgaaa gaggcctata taagtcttta aaaggagcat
     2941 gcaaactcaa gttatgtgga gttctaggac ttagacttat ggatggaaca tgggtctcga
     3001 tgcaaacatc aaatgaaacc aaatggtgcc ctcccgataa gttggtgaac ctgcacgact
     3061 ttcgctcaga cgaaattgag caccttgttg tagaggagtt ggtcaggaag agagaggagt
     3121 gtctggatgc actagagtcc atcatgacaa ccaagtcagt gagtttcaga cgtctcagtc
     3181 atttaagaaa acttgtccct gggtttggaa aagcatatac catattcaac aagaccttga
     3241 tggaagccga tgctcactac aagtcagtca gaacttggaa tgagatcctc ccttcaaaag
     3301 ggtgtttaag agttgggggg aggtgtcatc ctcatgtgaa cggggtgttt ttcaatggta
     3361 taatattagg acctgacggc aatgtcttaa tcccagagat gcaatcatcc ctcctccagc
     3421 aacatatgga gttgttggaa tcctcggtta tcccccttgt gcaccccctg gcagacccgt
     3481 ctaccgtttt caaggacggt gacgaggctg aggattttgt tgaagttcac cttcccgatg
     3541 tgcacaatca ggtctcagga gttgacttgg gtctcccgaa ctgggggaag tatgtattac
     3601 tgagtgcagg ggccctgact gccttgatgt tgataatttt cctgatgaca tgttgtagaa
     3661 gagtcaatcg atcagaacct acgcaacaca atctcagagg gacagggagg gaggtgtcag
     3721 tcactcccca aagcgggaag atcatatctt catgggaatc acacaagagt gggggtgaga
     3781 ccagactgta aggcgcgtcg agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3841 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3901 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3961 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4021 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4081 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4141 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4201 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4261 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4321 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4381 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4441 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4501 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4561 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4621 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4681 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4741 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4801 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4861 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4921 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4981 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5041 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5101 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5161 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5221 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5281 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5341 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5401 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5461 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5521 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5581 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5641 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5701 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5761 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5821 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5881 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5941 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6001 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6061 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6121 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6181 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6241 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6541 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
