LOCUS       sequence-60329-e        5904 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 5904)
  AUTHORS   Shigetomi E, Bushong EA, Haustein MD, Tong X, Jackson-Weaver O, 
            Kracun S, Xu J, Sofroniew MV, Ellisman MH, Khakh BS
  TITLE     Imaging calcium microdomains within entire astrocyte territories and
            endfeet with GCaMPs expressed using adeno-associated viruses.
  JOURNAL   J Gen Physiol. 2013 Apr 15.
  PUBMED    23589582
REFERENCE   2  (bases 1 to 5904)
  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 Gen 
            Physiol. 2013 Apr 15."
COMMENT     Sequence Label: pZac2.1 gfaABC1D-tdTomato
FEATURES             Location/Qualifiers
     source          1..5904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1451..1460
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1457..2887
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    2929..3050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2994..3013
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3212..3341
                     /label=AAV2 ITR
     primer_bind     complement(3360..3377)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3360..3376)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3369..3391)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3518..3973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3605..3624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3815..3836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3999..4103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4104..4172,4173..4964)
                     /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"
     rep_origin      5135..5723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5624..5643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     1441 tcgagaattc gccaccatgg tgagcaaggg cgaggaggtc atcaaagagt tcatgcgctt
     1501 caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     1561 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
     1621 gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa
     1681 gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga
     1741 gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca
     1801 ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc
     1861 cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga
     1921 cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccactacct
     1981 ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg gctactacta
     2041 cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2101 cgagcgctcc gagggccgcc accacctgtt cctggggcat ggcaccggca gcaccggcag
     2161 cggcagctcc ggcaccgcct cctccgagga caacaacatg gccgtcatca aagagttcat
     2221 gcgcttcaag gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga
     2281 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     2341 ccccctgccc ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta
     2401 cgtgaagcac cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa
     2461 gtgggagcgc gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc
     2521 cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga
     2581 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc
     2641 ccgcgacggc gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca
     2701 ctacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta
     2761 ctactacgtg gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2821 acagtacgag cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta
     2881 caagtagatt ctagagtcga cccgggcggc cgcttcgagc agacatgata agatacattg
     2941 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3001 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3061 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     3121 aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat
     3181 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     3241 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3301 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca
     3361 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3421 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3481 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3961 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4021 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4081 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4141 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4201 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4261 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4321 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4381 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4441 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4501 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4561 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4621 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4681 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4741 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4801 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4861 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4921 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5881 agtcagtgag cgaggaagcg gaag
//
