LOCUS       sequence-244970-        6720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human AQP4 (isoform M23) as an EmGFP fusion protein in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6720)
  AUTHORS   Mader S, Gredler V, Schanda K, Rostasy K, Dujmovic I, Pfaller K, 
            Lutterotti A, Jarius S, Di Pauli F, Kuenz B, Ehling R, Hegen H, 
            Deisenhammer F, Aboul-Enein F, Storch MK, Koson P, Drulovic J, 
            Kristoferitsch W, Berger T, Reindl M
  TITLE     Complement activating antibodies to myelin oligodendrocyte 
            glycoprotein in neuromyelitis optica and related disorders.
  JOURNAL   J Neuroinflammation. 2011 Dec 28;8:184. doi: 
            10.1186/1742-2094-8-184.
  PUBMED    22204662
REFERENCE   2  (bases 1 to 6720)
  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 
            Neuroinflammation."; date: "2011-12-28"; volume: "8:184. doi"; 
            pages: " 10.1186/1742-2094-8-184"
COMMENT     Sequence Label: pcDNA6.2 EmGFP hAQP4(M23)
FEATURES             Location/Qualifiers
     source          1..6720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    681..705
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1639..1663)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1677..1679,1680..1682,1683..2396)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1722..1743)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1983..2002)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2330..2351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2424..2443)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2468..2516
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2718..3146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2805..2824)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3015..3036
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3155..3175)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3160..3489
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3340..3475
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3402..3421
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3537..3584
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3603..4001
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4159..4280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4196..4215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4250..4269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4329..4345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4329..4345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4342..4364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4353..4369
                     /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(4377..4383,4384..4401,4402..4407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4422..4443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4560..4577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4731..5319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4811..4830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5490..6281,6282..6350))
                     /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     6113..6132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6351..6455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6530..6549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag ctgagcatca acaagtttgt acaaaaaagc aggctccgaa ttcgccctta
      721 tcatggtggc tttcaaaggg gtctggactc aagctttctg gaaagcagtc acagcggaat
      781 ttctggccat gcttattttt gttctcctca gcctgggatc caccatcaac tggggtggaa
      841 cagaaaagcc tttaccggtc gacatggttc tcatctccct ttgctttgga ctcagcattg
      901 caaccatggt gcagtgcttt ggccatatca gcggtggcca catcaaccct gcagtgactg
      961 tggccatggt gtgcaccagg aagatcagca tcgccaagtc tgtcttctac atcgcagccc
     1021 agtgcctggg ggccatcatt ggagcaggaa tcctctatct ggtcacacct cccagtgtgg
     1081 tgggaggcct gggagtcacc atggttcatg gaaatcttac cgctggtcat ggtctcctgg
     1141 ttgagttgat aatcacattt caattggtgt ttactatctt tgccagctgt gattccaaac
     1201 ggactgacgt cactggctca atagctttag caattggatt ttctgttgca attggacatt
     1261 tatttgcaat caattatact ggtgccagca tgaatcccgc ccgatccttt ggacctgcag
     1321 ttatcatggg aaattgggaa aaccattgga tatattgggt tgggcccatc ataggagctg
     1381 tcctcgctgg tggcctttat gagtatgtct tctgtccaga tgttgaattc aaacgtcgtt
     1441 ttaaagaagc cttcagcaaa gctgcccagc aaacaaaagg aagctacatg gaggtggagg
     1501 acaacaggag tcaggtagag acggatgacc tgattctaaa acctggagtg gtgcatgtga
     1561 ttgacgttga ccggggagag gagaagaagg ggaaagacca atctggagag gtattgtctt
     1621 cagtaaaggg cgaattcgac ccagctttct tgtacaaagt ggttgatgct gttaacatgg
     1681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1801 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1861 tgaccacctt cacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2101 tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca
     2161 tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2281 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatgat
     2401 aagtttaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     2461 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     2521 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     2581 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     2641 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     2701 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3061 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3121 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3241 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3361 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3421 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3481 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     3541 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     3601 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     3661 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     3721 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     3781 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     3841 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     3901 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     3961 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacttcgt ggccgaggag
     4021 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4081 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4141 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4201 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4261 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4321 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4381 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4441 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4501 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4561 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4621 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4681 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4741 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4801 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4861 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4921 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4981 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5041 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5101 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5161 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5221 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5281 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5341 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5401 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5461 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5521 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5581 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5641 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5701 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5761 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5821 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5881 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5941 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6001 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6061 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6121 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6181 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6241 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6301 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6481 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6541 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6601 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6661 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
