LOCUS       sequence-64388-e        7298 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 7298)
  TITLE     Fritjof Helmchen Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7298)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-EF1a-YCNano140
FEATURES             Location/Qualifiers
     source          1..7298
                     /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        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(1433..1435,1436..1438,1439..1951)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             2579..2656
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2663..3397
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     misc_feature    3425..4013
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3478..3498)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3896..3907)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4045..4521
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4244..4263)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4561..4701
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4561..4690
                     /label=AAV2 ITR
     rep_origin      4776..5231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4863..4882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5073..5094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5248..5267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5367..5389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5427..5445)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5513..5617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5618..5686,5687..6478)
                     /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(5836..5855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6649..7237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7138..7157
                     /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 cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ccatggtgag
     1441 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1501 aaacggccac aggttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1561 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1621 caccctgacc tggggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1681 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgtaccatct tcttcaagga
     1741 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1801 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1861 gtacaactac atcagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1921 ggcccacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1981 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2041 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2101 gttcgtgacc gccgcccgca tgcatgacca actgacagaa gagcagattg cagagttcaa
     2161 agaagccttc tcattattcg acaaggatgg ggacggcacc atcaccacaa aggaacttgg
     2221 caccgttatg aggtcgcttg gacaaaaccc aacggaagca gaattgcagg atatgatcaa
     2281 tgaagtcgat gctgatggca atggaacgat ttactttcct gaatttctta ctatgatggc
     2341 tagaaaaatg aaggacacag acagcgaaga ggaaatccga gaagcattcc gtgtttttga
     2401 caaggatggg aacggctaca tcagcgctgc tcagttacgt cacgtcatga caaacctcgg
     2461 ggagaagtta acagatgaag aagttgatga aatgataagg gaagcagata tcgatggtga
     2521 tggccaagta aactatgaag agtttgtaca aatgatgaca gcaaaggggg gcgggtccaa
     2581 gaggcgctgg aagaaaaact tcattgccgt cagcgctgcc aaccggttca agaagatctc
     2641 cagctccggg gcactggagc tcatggacgg cggcgtgcag ctcgccgacc actaccagca
     2701 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     2761 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2821 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtggca gcggtggcat
     2881 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2941 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3001 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3061 cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     3121 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3181 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3241 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3301 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     3361 catcaaggcc aacttcaaga tccgccacaa catcgagtaa gaattcgata tcaagcttat
     3421 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3481 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3541 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3601 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3661 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3721 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3781 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3841 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3901 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3961 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     4021 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     4081 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     4141 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     4201 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     4261 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     4321 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     4381 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     4441 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4501 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     4561 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4621 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4681 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4741 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4801 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4861 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4921 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4981 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5041 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5101 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     5161 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5221 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5281 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5341 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5401 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5461 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5521 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5581 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5641 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5701 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5761 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5821 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5881 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5941 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6001 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6061 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6121 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6181 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6241 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6301 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6361 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6421 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6481 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6541 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6601 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6661 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6721 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6781 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6841 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6901 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6961 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7021 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7081 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7141 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7201 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7261 acggttcctg gccttttgct ggccttttgc tcacatgt
//
