LOCUS       sequence-405431-        5032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For barcoded retrograde labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5032)
  AUTHORS   Xu P, Peng J, Yuan T, Chen Z, He H, Wu Z, Li T, Li X, Wang L, Gao L,
            Yan J, Wei W, Li CT, Luo ZG, Chen Y
  TITLE     High-throughput mapping of single-neuron projection and molecular 
            features by retrograde barcoded labeling.
  JOURNAL   Elife. 2024 Feb 23;13:e85419. doi: 10.7554/eLife.85419.
  PUBMED    38390967
REFERENCE   2  (bases 1 to 5032)
  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: "Elife."; 
            date: "2024-02-23"; volume: "13:e85419. doi"; pages: " 
            10.7554/eLife.85419"
COMMENT     Sequence Label: pAAV-CAG-EGFPnls barcode-206-SV40 polyA
FEATURES             Location/Qualifiers
     source          1..5032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     primer_bind     1048..1070
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1132..1151
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1193..1202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1199..1201,1202..1204,1205..1915)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1199..1201,1202..1204,1205..1915)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1244..1265)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1505..1524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1852..1873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1916..1963
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(2023..2040)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2029..2253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2762..3350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2842..2861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3521..4312,4313..4381))
                     /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     4144..4163
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4382..4486)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4512..4967)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4649..4670)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4861..4880
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctagga agagtaccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      301 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      361 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      421 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      481 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      541 agcgatgggg gcgggggggg gggggggksk kgydmnngkg gggggngggg gggggggggg
      601 gggggggggg gggsgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag
      661 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
      721 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      781 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      841 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      901 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      961 gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg ggggacggct
     1021 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     1081 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     1141 tggttattgt gctgtctcat cattttggca aagaattgga tccgtcgacg ccgccaccat
     1201 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1261 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1321 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1381 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1441 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1501 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1561 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1621 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1681 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1741 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1801 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1861 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagaagcg
     1921 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaataatgag aattcgcgta
     1981 agtctccttg ggatccaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     2041 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2101 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2161 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2221 gcaggcatgc tggggatgcg gtgggctcta tgggttaaca acaacaattg cattcatttt
     2281 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2341 tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     2401 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2461 tcgctcgctc actgaggccg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2521 cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2581 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2641 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2701 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2761 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2821 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2881 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2941 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3001 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3061 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3121 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3181 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3241 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3301 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3361 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3421 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3481 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3601 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3661 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3721 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3781 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3841 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3901 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3961 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4021 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4081 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4141 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4201 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4261 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4321 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4381 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4441 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4501 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     4561 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4621 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4681 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4741 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4801 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4861 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4921 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     4981 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat ta
//
