LOCUS       sequence-348849-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cx43 fused with GFP at the C-terminus in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Soto JS, Jami-Alahmadi Y, Chacon J, Moye SL, Diaz-Castro B, 
            Wohlschlegel JA, Khakh BS
  TITLE     Astrocyte-neuron subproteomes and obsessive-compulsive disorder 
            mechanisms.
  JOURNAL   Nature. 2023 Apr;616(7958):764-773. doi: 10.1038/s41586-023-05927-7.
            Epub 2023 Apr 12.
  PUBMED    37046092
REFERENCE   2  (bases 1 to 6991)
  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: 
            "10.1038/s41586-023-05927-7"; journalName: "Nature"; date: 
            "2023-04"; volume: "616"; issue: "7958"; pages: "764-773"
COMMENT     Sequence Label: PZac2.1 gfaABC1D-Cx43-GFP
FEATURES             Location/Qualifiers
     source          1..6991
                     /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      2619..2628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2625..2627,2628..2630,2631..3344)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2625..2627,2628..2630,2631..3341)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2670..2691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2931..2950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3278..3299
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3379..3967
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3432..3452)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3850..3861)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4016..4137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4027..4046)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4081..4100
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4299..4428
                     /label=AAV2 ITR
     primer_bind     complement(4447..4464)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4447..4463)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4456..4478)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4605..5060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4692..4711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4902..4923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5086..5190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5191..5259,5260..6051)
                     /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      6222..6810
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6711..6730
                     /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 tcgagctcaa gcttcgaatt atgggtgact ggagtgcctt ggggaagctt ctggacaagg
     1501 tccaagccta ctccaccgct ggagggaagg tgtggctgtc agtgctcttc atattcagaa
     1561 tcctgctcct ggggacagct gttgagtcag cttggggtga tgaacagtct gcctttcgct
     1621 gtaacactca acaacctggc tgcgaaaacg tctgctatga caagtccttc cccatctctc
     1681 acgtgcgctt ctgggtcctt cagatcatat tcgtgtctgt gcccaccctc ctgtacttgg
     1741 cccatgtgtt ctatgtgatg aggaaggaag agaagctaaa caagaaagaa gaggagctca
     1801 aagtggccca gactgacggg gtcaacgtgg agatgcacct gaagcagatt gaaatcaaga
     1861 agttcaagta cgggattgaa gagcacggca aggtgaaaat gaggggcggc ttgctgagaa
     1921 cctacatcat cagcatcctc ttcaagtctg tcttcgaggt ggccttcctg ctcatccagt
     1981 ggtacatcta tgggttcagc ttgagcgcgg tctacacctg caagagagat ccctgccccc
     2041 accaggtaga ctgcttcctc tcacgtccca cggagaaaac catcttcatc atcttcatgc
     2101 tggtggtgtc cttggtgtct ctcgctttga acatcattga gctcttctac gtcttcttca
     2161 aaggcgttaa ggatcgcgtg aagggaagaa gcgatcctta ccacgccacc actggcccac
     2221 tgagcccatc aaaagactgc ggatctccaa aatacgccta cttcaatggc tgctcctcac
     2281 caacggctcc actctcgcct atgtctcctc ctgggtacaa gctggttact ggtgacagaa
     2341 acaattcctc gtgccgcaat tacaacaagc aagctagcga gcaaaactgg gcgaactaca
     2401 gcgcagagca aaatcgcatg gggcaggccg gaagcaccat ctccaactcg cacgcccagc
     2461 cgttcgattt ccccgacgac aaccagaatg ccaaaaaagt tgctgctgga catgaactcc
     2521 agccattagc catcgtggac caacgacctt ccagcagagc cagcagccgc gccagcagca
     2581 ggcctcggcc tgatgacctg gagattgatc caccggtcgc caccatggtg agcaagggag
     2641 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2701 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2761 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2821 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2881 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2941 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3001 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3061 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3121 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3181 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3241 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3301 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgaccggt
     3361 atcatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3421 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3481 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3541 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3601 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3661 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3721 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3781 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     3841 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3901 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3961 tccccgcatc gatacctcta gagtcgaccc gggcggccgc ttcgagcaga catgataaga
     4021 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     4081 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4141 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     4201 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     4261 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     4321 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4381 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc ttaattaacc
     4441 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4501 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4561 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4621 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4681 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4741 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4801 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4861 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4921 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4981 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5041 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5101 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5161 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5221 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5281 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5341 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5401 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5461 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5521 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5581 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5641 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5701 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5761 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5821 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5881 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5941 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6001 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6061 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6121 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6181 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6241 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6301 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6361 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6421 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6481 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6541 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6601 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6661 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6721 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6781 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6841 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6901 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6961 cgcagcgagt cagtgagcga ggaagcggaa g
//
