LOCUS       sequence-494690-        7632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SynBar protein and RNA construct used in HEK and neuron culture 
            experiment..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7632)
  AUTHORS   Chen D, Isakova A, Wan ZJ, Wagner MJ, Wu Y, Zhao BS
  TITLE     Connectome-seq: High-throughput Mapping of Neuronal Connectivity at 
            Single-Synapse Resolution via Barcode Sequencing.
  JOURNAL   bioRxiv [Preprint]. 2025 May 26:2025.02.13.638129. doi: 
            10.1101/2025.02.13.638129.
  PUBMED    41030982
REFERENCE   2  (bases 1 to 7632)
  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: "bioRxiv 
            [Preprint]. 2025 May 26:2025.02.13.638129. doi: 
            10.1101/2025.02.13.638129."
COMMENT     Sequence Label: hSyn-PostSynBar-hSyn-PostRNA short
FEATURES             Location/Qualifiers
     source          1..7632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             648..692
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             693..716
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1208..1217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2547..2594
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             3039..3065
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3072..3098
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3306..3371
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     misc_feature    3399..3987
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3452..3472)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3870..3881)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4042..4489
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    4550..4568
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    4574..4592
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     primer_bind     complement(4653..4670)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4659..4866
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4895..5035
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4895..5024
                     /label=AAV2 ITR
     rep_origin      5110..5565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5197..5216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5407..5428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5582..5601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5701..5723
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5761..5779)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5847..5951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5952..6020,6021..6812)
                     /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(6170..6189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6983..7571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7472..7491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg tctgcacttc
      661 tgatcctagc tcttgttgga gctgcagttg ctgactacaa agacgatgac gacaagcttg
      721 cggccgcgaa ttcatcgata gatctgcaaa agttggatga tgtagaccca ttggttacta
      781 ctaactttgg caaaataagg ggaattaaga aagaactcaa taatgaaatt ttggggcctg
      841 tcattcagtt tcttggggtt ccatatgctg ctccaccaac aggagaacat cgatttcagc
      901 ctccagaacc accatctccc tggtctgata tccggaatgc cactcagttt gctcctgtat
      961 gtccccagaa tatcattgat ggcagattgc ctgaagttat gcttcctgtg tggttcacta
     1021 ataacttgga tgtggtttcc tcatatgtgc aagaccagag tgaagactgt ctatacttaa
     1081 acatctatgt cccaactgaa gatgtaaaaa gaatatccaa ggaatgtgcc agaaaacccg
     1141 gcaagaaaat atgtagaaaa ggagatattc gggacagtgg gggtcccaaa ccagtgatgg
     1201 tgtacatcca tggtggctct tacatggaag gtactggaaa tctatatgat gggagtgtgt
     1261 tggcaagcta cggtaatgtg atcgtcatca cagtcaacta tcggcttggg gtacttggct
     1321 tcttgagcac aggggatcag gctgccaaag gaaactacgg gctccttgac ctcatccagg
     1381 ccctcagatg gaccagcgag aacattgggt tctttggtgg tgaccccttg cgaatcaccg
     1441 tgtttggatc aggcgctggg ggttcatgtg tcaatctgct gactttatcc cattattctg
     1501 aaaaaggact ttttcaacga gcaatagctc agagtggaac agccctttcc agctgggctg
     1561 ttagtttcca gcctgcaaaa tacgctagaa ttctagccac aaaagttggc tgcaatgtgt
     1621 cagatacagt agagttagta gaatgcctgc agaagaagcc ttacaaagaa cttgttgatc
     1681 aagatgttca accagcccga taccacatag cctttggacc tgtgatcgat ggtgatgtaa
     1741 taccagatga ccctcagata ctgatggaac aaggagagtt cctcaattat gatatcatgt
     1801 taggagttaa ccaaggggaa gggttgaagt ttgtggaaaa catagtagat agtgatgatg
     1861 gtgtatcagc cagtgatttt gactttgctg tctctaattt tgttgataat ttatatggat
     1921 atcctgaagg caaagatgtt ttgagagaaa ccattaaatt catgtatact gactgggctg
     1981 atcgccataa ccctgaaact agaaggaaga cattgttggc tttgtttaca gaccatcagt
     2041 gggtagcacc cgctgtggct acagcagacc ttcactcgaa ctttggctca cctacatact
     2101 tctatgcctt ttatcatcat tgccaaactg accaagttcc agcttgggct gatgcagctc
     2161 atggggatga ggttccctat gtattgggaa tccccatgat tggccctaca gagttatttc
     2221 cttgcaattt ctccaagaat gatgtgatgt tgagtgctgt agtaatgaca tactggacga
     2281 attttgctaa aaccggtgac ccaaatcaac cagttcctca agacacaaaa ttcattcata
     2341 ccaaacccaa ccgctttgaa gaagtagcat ggaccagata ttcccagaaa gaccaacttt
     2401 atctccatat tggattaaaa ccgagagtta aagaacatta cagagccaat aaggtaaatc
     2461 tctggttgga gctggtacct catctgcata atctcaatga catttctcag acgcgtggag
     2521 gttcaggtgg aggttcagga ggaggaagag atcacatggt tttgcacgaa tacgtgaatg
     2581 ctgcaggaat tacaggtgga ggttctggag gtggttctgg aggtacgcgt tatacctcga
     2641 caacaactaa agtgccatca acagacatca ctctcagacc tacaaggaaa aattccactc
     2701 cagtcacatc agcctttccc actgccaagc aggatgatcc caaacaacaa ccaagtccct
     2761 tctcggtaga tcaaagggac tactccacag agctaagtgt cacaatcgca gttggagcct
     2821 ctctgctgtt tctgaacatc ttggcttttg cagccctata ctacaagaaa gacaagagga
     2881 gacatgatgt ccaccggagg tgcagccctc agcgcaccac gaccaacgac ctaacccatg
     2941 ctccagaaga ggaaattatg tctctccaaa tgaagcacac tgacttggat cacgagtgtg
     3001 agtccatcca tccacatgag gtggttcttc ggaccgccta cccatacgat gttccagatt
     3061 acgctgctag ctatccttat gacgtgcctg actatgcccg gaccgcctgc cccccagatt
     3121 atactcttgc tatgaggagg tcaccggatg atgttccttt aatgacacct aacaccatca
     3181 caatgattcc caacactata ccagggattc agcccttaca tacattcaac acatttactg
     3241 gaggacagaa taatacactg ccccaccccc accctcaccc ccattcacat tcaacaacca
     3301 gggtaatgga cgcacaaaca cgacgacgtg agcgtcgcgc tgagaaacaa gctcaatgga
     3361 aagctgcaaa ctaggaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     3421 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3481 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3541 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3601 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3661 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3721 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3781 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     3841 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3901 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3961 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgcg agtgtctaga
     4021 ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg
     4081 gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca
     4141 aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc
     4201 actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg
     4261 ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc
     4321 cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg
     4381 cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc
     4441 tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgagaacgta
     4501 cgttccctaa gtccaactac taaactgggc taagtccaac tactaaactg ggccctgaag
     4561 aagggcccat atagggccct gaagaagggc cctatcgagg atattatctc gagatgcatc
     4621 tagtacgcaa ttggctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4681 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4741 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4801 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4861 tggggaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4921 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4981 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     5041 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     5101 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5161 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     5221 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5281 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5341 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5401 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     5461 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5521 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5581 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5641 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5701 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5761 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5821 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5881 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5941 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6001 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6061 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6121 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6181 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6241 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6301 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6361 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6421 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6481 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6541 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6601 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6661 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6721 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6781 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6841 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6901 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6961 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7021 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7081 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7141 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7201 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7261 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7321 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7381 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7441 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7501 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7561 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7621 ttgctcacat gt
//
