LOCUS       sequence-494692-        7463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The set of SynBar constructs used in the optimized connectome-seq 
            experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7463)
  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 7463)
  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 barcode landing pad
FEATURES             Location/Qualifiers
     source          1..7463
                     /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    4479..4497
                     /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    4503..4521
                     /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    4565..4686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4602..4621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4656..4675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4726..4866
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4726..4855
                     /label=AAV2 ITR
     rep_origin      4941..5396
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5028..5047)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5238..5259
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5413..5432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5532..5554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5592..5610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5678..5782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5783..5851,5852..6643)
                     /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(6001..6020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6814..7402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7303..7322
                     /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 caaccacaac 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 caaattgcgt 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 gcaattggag accggagaag ccctttgcca tcgccaagga gtagggcaca
     4261 gggacatctt tctttgagtg accgtctgtg caggccctgt agtccgccac agggctctga
     4321 gctgcactgg ccccggtgct ggcatctggt ggagcggacc cactcccctc acattccaca
     4381 ggcccatgga ctcactttgg tctcatgcat cctctttttc cggctggaac cttccctaag
     4441 tccaactact aaactgggct aagtccaact actaaactgg gccctgaaga agggcccata
     4501 tagggccctg aagaagggcc ctatcgagga tattatctcg aggctttaag gccggtccta
     4561 gcaaaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4621 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4681 atcttacatg ctggggaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4741 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4801 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4861 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4921 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4981 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     5041 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5101 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     5161 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5221 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     5281 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     5341 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     5401 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5461 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5521 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5581 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5641 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5761 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5821 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5881 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5941 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6001 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6061 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6121 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6181 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6241 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6301 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6361 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6421 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6481 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     6541 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6601 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6661 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6721 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6781 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6841 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6901 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6961 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7141 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7201 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7321 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7381 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7441 ttgctggcct tttgctcaca tgt
//
