LOCUS       sequence-494689-        7254 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 7254)
  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 7254)
  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-PreSynBar-hSyn-PreRNA short
FEATURES             Location/Qualifiers
     source          1..7254
                     /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..734
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2580..2589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2586..2651
                     /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    2890..3478
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2943..2963)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3361..3372)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3520..3537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3526..3733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3764..4211
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    4272..4290
                     /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    4296..4314
                     /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)"
     polyA_signal    4356..4477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4393..4412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4447..4466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4517..4657
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4517..4646
                     /label=AAV2 ITR
     rep_origin      4732..5187
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4819..4838)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5029..5050
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5204..5223)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5323..5345
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5383..5401)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5469..5573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5574..5642,5643..6434)
                     /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(5792..5811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6605..7193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7094..7113
                     /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 ctggcaagcc gatccctaac cctctgctgg
      721 gcctggacag cactcttgcg gccgcggcat ccagtttggg agcgcaccac atccaccatt
      781 tccatggcag cagcaagcat cattcagtgc ctattgcaat ctacaggtca ccagcatcct
      841 tgcgaggcgg acacgctggg acgacgtata tctttagcaa aggtggtgga caaattacat
      901 ataagtggcc tcctaatgac cgacccagta cacgagcaga caggctggcc atcgggttta
      961 gcactgttca gaaggaagca gtgttggtgc gtgtggacag ttcctcgggc ctgggtgact
     1021 accttgagct gcacatacac caaggaaaaa ttggagttaa atttaatgtt gggacagatg
     1081 acatcgccat tgaggagtcc aatgcaatca ttaatgatgg aaaataccat gtagtgcgtt
     1141 tcacgaggag tggtggcaat gccacgttac aggtggacag ctggccagtc atcgaacgat
     1201 accctgcatc cggaggaggt tcaggtggag gttcaggagg aggaatgggt ggaacaagca
     1261 tgagcaaagg tgaggaactg ttcaccggcg tcgtacctat tcttgtggag ctggacgggg
     1321 acgtcaatgg acacaaattc tcagttcgcg gcgagggcga gggggatgct accatcggta
     1381 agctgaccct caaattcatc tgcactacgg gcaagctgcc ggttccctgg cctactctgg
     1441 tcactacact gacttacggg gtccaatgct tctccagata tcccgaccac atgaagcggc
     1501 acgacttctt taaaagcgcc atgccggagg ggtacgtgca ggaacgaacc atttccttca
     1561 aagatgatgg taagtataag accagagctg tcgtcaagtt tgagggcgac accctcgtga
     1621 accgcatcga acttaaggga accgatttta aggaagacgg taacatcctg ggccacaagc
     1681 tggagtacaa tttcaactca cataatgtgt acattacggc agacaagcag aaaaacggga
     1741 tcaaagccaa ttttacagta cggcacaatg tcgaggacgg cagcgtccag ctggcagatc
     1801 attaccagca aaacacgcct atcggagatg ggccggtgct gcttccagat aaccattacc
     1861 tttccacaca gaccgtgctg agcaaggacc ctaacgagaa gggtggaggt tctggaggtg
     1921 gttctggagg ttccggaggg cgtcagctca caatcttcaa tagccaagca accataataa
     1981 ttggcgggaa agagcagggc cagcccttcc agggccagct ctctgggctt tactacaatg
     2041 gcttgaaagt tctgaatatg gcggcagaga acgatgccaa catcgccata gtgggaaatg
     2101 tgaggctggt cggtgaagtg ccttcctcta tgacaactga gtcgacagcc actgccatgc
     2161 agtccgagat gtccacctca atcatggaga ccaccacaac cctggctacc agcacagctc
     2221 gacgaggaaa gccccccaca aaggagccta tcagccagac cacggatgat atccttgtgg
     2281 cctcggcaga gtgtcccagc gacgatgagg acattgaccc ctgtgagccg agctcaggtg
     2341 ggttagccaa ccccacccga gtgggcggac gcgaaccata cccaggctcg gcagaggtga
     2401 tccgggagtc tagcagtacc actggcatgg tggtggggat tgtcgcagca gcagctctgt
     2461 gcatcctcat cctcctctat gccatgtaca agtacaggaa ccgggatgaa ggttcaagcg
     2521 ctccaccgct cgacggagcc ggagctggcg ctggagctgg agccggagct ggcgggctag
     2581 ccaccatgga cgcacaaaca cgacgacgtg agcgtcgcgc tgagaaacaa gctcaatgga
     2641 aagctgcaaa cctcgagcca ccgctcgacg gagccggagc tggcgctgga gctggagccg
     2701 gagctggcgg tctagccacc ccaccgagcg cttatcacgt ggatgagagt cgaaactaca
     2761 tcagtaactc agcacagtcc aatggggctg tggtcaagga gaagcagccc agcagtgcga
     2821 aaagcgccaa caaaaacaag aagaacaagg ataaggagta ttatgtctga cgatatcaag
     2881 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2941 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3001 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3061 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3121 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3181 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3241 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3301 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3361 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3421 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3481 cgataccgag cgctgctcga gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3721 ggcatgctgg ggagtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
     3781 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
     3841 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
     3901 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
     3961 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
     4021 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
     4081 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
     4141 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
     4201 tgagagcgca gtcgagaacg tacgttccct aagtccaact actaaactgg gctaagtcca
     4261 actactaaac tgggccctga agaagggccc atatagggcc ctgaagaagg gccctatcga
     4321 ggatattatc tcgagcaatt gctagtacga tgcataactt gtttattgca gcttataatg
     4381 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4441 ctagttgtgg tttgtccaaa ctcatcaatg tatcttacat gctggggagg taaccacgtg
     4501 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4561 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4621 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4681 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4741 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4801 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4861 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4921 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4981 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5041 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     5101 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5161 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     5221 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5281 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5341 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5401 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5461 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5521 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5581 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5641 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5701 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5761 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5821 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5881 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5941 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6001 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6061 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6121 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6181 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6241 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     6301 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6361 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6421 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6481 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6541 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7201 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
