LOCUS       sequence-376443-        4978 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 4978)
  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 4978)
  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-EGFP barcode-7-SV40 polyA
FEATURES             Location/Qualifiers
     source          1..4978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     881..903
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     965..984
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1030..1039
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1036..1038,1039..1041,1042..1755)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1036..1038,1039..1041,1042..1752)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1081..1102)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1342..1361)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1689..1710
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1825..1842)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1831..2055
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2220..2349
                     /label=AAV2 ITR
     rep_origin      complement(2586..3174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2666..2685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3345..4136,4137..4205))
                     /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     3968..3987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4206..4310)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4336..4791)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4473..4494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4685..4704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(4863..4978,1..14)
                     /label=AAV2 ITR
ORIGIN
        1 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
       61 ctctaggaag agtaccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      361 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      421 cgatgggggc gggggggggg ggggggcgcg cgccrggggg gggggggggg gggggggggg
      481 ggggsggggs grggsggrgr ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      541 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      601 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      661 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      721 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      781 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg
      841 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      901 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
      961 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccccgg
     1021 gtaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1081 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1141 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1201 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1261 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1321 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1381 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1441 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1501 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1561 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1621 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1681 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1741 gagctgtaca agtaaagcgg ccgcgactct agaattctgt aaggctactt ggggatccag
     1801 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1861 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1921 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1981 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2041 cggtgggctc tatgggttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2101 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     2161 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     2221 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2281 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2341 agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3481 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4321 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4381 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4441 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4501 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4561 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4621 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4681 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4741 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     4801 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4861 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     4921 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccat
//
