LOCUS       sequence-308125-        5573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Super-bright fluorescence protein for tracking and visualization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5573)
  TITLE     Chronos with red nucleus-targeted fluorescence for excitatory 
            neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5573)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-EF1-fDIO-mNeon-WPRE-PolyA
FEATURES             Location/Qualifiers
     source          1..5573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        157..368
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             381..649
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    695..728
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(824..1531)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     protein_bind    complement(1538..1571)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    1680..2268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1733..1753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2151..2162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2300..2776
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2499..2518)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2816..2956
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2816..2945
                     /label=AAV2 ITR
     rep_origin      3031..3486
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3118..3137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3328..3349
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3503..3522)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3622..3644
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3682..3700)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3768..3872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3873..3941,3942..4733)
                     /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(4091..4110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4904..5492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5393..5412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5554..5573,1..110)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
       61 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac
      121 gcgtaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      181 tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg
      241 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      301 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      361 gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct
      421 gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct
      481 gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg
      541 cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg
      601 ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac
      661 cggcgcctac ggatcctcta gagtcgactc cggagaagtt cctattctct agaaagtata
      721 ggaacttcgc agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaga
      781 agttcctatt cttcaaaagg tataggaact tcggcgcgcc taattacttg tacagctcgt
      841 ccatgcccat cacatcggta aaggcctttt gccactcctt gaagttgagc tcggtcttgg
      901 agtgcttgag ctccgtctta cggaacacgt acatcggctg gttcttcaga tagttagccg
      961 ccattggctt ggcaaaggtg taggtggtcc gcgcagtgct ccggtagcgc ttgccatttc
     1021 cagtggtgta actccactta aaggtactga tgatggtttt gtcgttgggg taagtcttct
     1081 tcgacctgca ccagtccgca gcggtcagcg agttggtcat cacaggaccg tcagcaggga
     1141 aaccagtccc cttcacctgg gcctctcctt tgatgtggct tccctcgtag gtgtagcggt
     1201 agttaacagt aagggaggca ccatcttcaa actgcattgt gcgatggact tggtatccag
     1261 agccatctac catagcggcc tggaaaggcg acatcccgtc agggtagggc aggtactgat
     1321 ggaagccata cccgatatga gggaccagaa tccaggggga gaactggagg tcgcccttgg
     1381 tggacttcag gtttaactcc tcataaccat catttggatt gccggtgccc tgacccacca
     1441 tgtcaaagtc cacaccgttg atggagccaa agatgtgtaa ctcatgtgtc gctgggagag
     1501 aggccatgtt atcctcctcg cccttgctca cgctagcgaa gttcctatac tttctagaga
     1561 ataggaactt cttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     1621 agaagttcct ataccttttg aagaatagga acttcgaatt cgatatcaag cttatcgata
     1681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     1741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     1801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     1861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     1921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     1981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2041 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2101 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     2281 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     2341 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     2401 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2461 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2521 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2581 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2641 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     2701 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     2761 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     2821 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2881 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2941 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3001 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3061 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     3121 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3181 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3241 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3301 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3361 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     3421 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3481 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3541 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3601 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3661 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3721 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3781 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3841 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3901 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3961 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4021 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4081 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4141 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4201 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4261 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4321 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4381 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4441 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4501 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4561 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4621 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4681 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4741 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4801 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4861 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4921 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4981 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5041 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5101 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5161 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5221 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5281 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5341 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5401 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5461 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5521 tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc gct
//
