LOCUS       sequence-480610-        6981 bp    DNA     circular SYN 25-JUL-2025
DEFINITION  Expresses synaptically localized dL5 protein fused to mNeonGreen, 
            under control of UAS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  AUTHORS   Sharma H, Robea MA, McGrory NH, Bazan DC, Burton EA, Burgess HA
  TITLE     Functional interrogation of neuronal connections by chemoptogenetic 
            presynaptic ablation.
  JOURNAL   bioRxiv [Preprint]. 2025 Apr 5:2025.04.04.647277. doi: 
            10.1101/2025.04.04.647277.
  PUBMED    40236196
REFERENCE   2  (bases 1 to 6981)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Apr 5:2025.04.04.647277. doi: 
            10.1101/2025.04.04.647277."
COMMENT     Sequence Label: UAS:syp-dL5-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..6981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     regulatory      1558..1567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2485..2494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2515..3234
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     primer_bind     complement(4585..4602)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4585..4601)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4594..4616)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4810..4829)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4929..4951
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4989..5007)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5075..5179
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5180..5248,5249..6040)
                     /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(5398..5417)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6211..6799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6700..6719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6953..6970
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atggtccctt tagccagtag cggttctagg cacgggccgt ccgggcggtg gcctggggcg
      301 gaaaactgaa ggggggcggc accggcggct cagcccttgt aatatattat atgcaccact
      361 attggtttac ttatgtcaca gtttgtaagt ttgtaacagc ctgaacctgg atcagcttag
      421 gcctccaagg cggagtactg tcctccgggc tggcggagta ctgtcctccg gcaaggtcgg
      481 agtactgtcc tccgacacta gaggtcggag tactgtcctc cgacgcaagg cggagtactg
      541 tcctccgggc tgcggagtac tgtcctccgg caaggtcgga gtactgtcct ccgacactag
      601 aggtcggagt actgtcctcc gacgcaaggt cggagtactg tcctccgaca ctagaggtcg
      661 gagtactgtc ctccgacgca aggtcggagt actgtcctcc gacactagag gtcggagtac
      721 tgtcctccga cgcaaggcgg agtactgtcc tccgggctgg cggagtactg tcctccggca
      781 agggtcgact ctagagggta tataatggat cccatcgcgt ctcagcctca ctttgagctc
      841 ctccacacga attcgatact aaggatcctt gtttgtgagt atgatgttaa atataagcaa
      901 tcgacgtttg ttttgtcgtt tgttaaatct agcaatgttt tatttcaatt aaagtgttaa
      961 ccggttcatt taaaggtgta tttctgaatg agctaacgta ttagctttta acgctagccg
     1021 gactgtttgg ttttgaggat caataaacca acgttttttt ctgatttatt ttgctttaaa
     1081 atataaaggt ataattcgtc gtcgagaatt ttgggttttg ttcgagttta gaaatttggc
     1141 ggcgtttttt tgtaccgcga aacagggcgg tggttcggtg agcttctcaa ttcatctgct
     1201 tgacaaacgg ttttcggcaa ctgtaaatgc gctatgatgc ttttattctt ttcagcgtgt
     1261 aactttgtta aatataattg actttccgac tatcgacgtt cgtttcatct ccgtgttatt
     1321 gtttaatggc cattttctgt ggtaaactgt cgccattttg ttttcagtcg tccgtttgtt
     1381 ccgcccttac tcacagctgt gtcagcatta gtgatgactg ttctcacctg gcctgtttgc
     1441 tttcattatt gatgaagagc tttctagaga attctgaagc tcgtctttgt cctgaattca
     1501 tctgtattca tttaaaaaaa aaaattaacc ttaatttttt tttcttctca gcgtaaagcc
     1561 accatggatg ttgccaacca gttggtcgcc actgggcata caatcgtcaa acagccatta
     1621 ggattcatca aaatcttaca atggttcttt gccatcttcg cgttcgccac ctgcggtggt
     1681 tactctggag tgtttcgtat gagcgtggag tgtgagaacc gctcagagag tgacctgaac
     1741 attgaggttg agtttgcgta cccgttcaga ctgcaccagg tatggtttga tgcccctacg
     1801 tgtaaaggat cagaacccga gcgactcttt ctggtgggag acaactcttc ctcagcagag
     1861 ttcttcgtca ccatcggcgt cttctccttc ctttattcaa tggcagccat ttctgtgtac
     1921 atcttcctgt tggagaaata ccgcgaggga aacaaaggag ctcaggctga tttcattgtg
     1981 acagcggttt tcaccttcat gtggctggtg agttcatcgg cgtgggctaa aggtctctca
     2041 gacgtgaaga gagccaccga ccctgatgac gttatcaacc tcatccctgc gtgcgaacga
     2101 gaggaaaacc gatgcaaaga gctgcacgaa ccggtggtgt ctggcctgaa cacatctgtg
     2161 gcattcgggt tctgtaatgt gatcctatgg ggaggcaacc tctggttcat cttcaaggaa
     2221 accggctgga tggcggcttt cgctggcacc tacatgccat ctggagagaa gcagccagct
     2281 ccagactcct tcggccaggg ctatggacag gaaggttatg gacaggacca gtatgccagc
     2341 tctcagggtg gatatcagcc tgactacggc cagcagggtg gatatgaagg cggtggatac
     2401 aatcaaggag tgtatggtca gggagagccc acatccttct ccaacgagat gtgtggagga
     2461 ggacccggga tccaccggcc ggtcgccacc atggaattcg gcctttctgg cctgcaggcc
     2521 gtcgttaccc aagaacctag tgttaccgtt agcccaggtg gtactgttat acttacttgt
     2581 ggaagtggta cgggtgccgt cacatctggt cattatgcaa attggtttca acaaaaacca
     2641 ggacaagctc caagagcttt gatttttgat actgataaga agtattcttg gaccccaggt
     2701 agattttctg gatctttgct gggagcaaag gcagctttga caatatcaga tgctcagcct
     2761 gaggacgaag ccgagtatta ctgttctctt agcgacgtgg atggctactt gtttggcggt
     2821 ggaacacaac tgacggttct gtccggtggt ggcggctctg gtggcggtgg cagcggcggt
     2881 ggtggttccg gaggcggcgg ttctcaggct gtggtgactc aggagccgtc agtgactgtg
     2941 tccccaggag ggacagtcat tctcacttgt ggctccggca ctggagctgt caccagtggt
     3001 cattatgcca actggttcca gcagaagcct ggccaagccc ccagggcact tatatttgac
     3061 accgacaaga agtattcctg gacccctggc cgattctcag gctccctcct tggggccaag
     3121 gctgccctga ccatctcgga tgcgcagcct gaagatgagg ctgagtatta ctgttcgctc
     3181 tccgacgttg acggttatct gttcggagga ggcacccagc tgaccgtcct ctccgcggcc
     3241 ggtctggccc atatgggagg cggtgggtct ggtggcggtg gcagcaccgg tatggtgagc
     3301 aagggagagg aggacaacat ggcttctctg cctgccaccc atgagctgca catctttgga
     3361 agcatcaatg gagtggactt tgacatggtg ggacagggaa ctggaaaccc aaatgatgga
     3421 tatgaggagc tgaacctgaa gagcaccaag ggagacctcc agttctctcc ttggattctg
     3481 gtgcctcaca ttggatatgg attccaccag tacctccctt accctgatgg aatgtctcct
     3541 ttccaggccg ctatggtgga tggatctgga taccaggtgc acagaaccat gcagtttgag
     3601 gatggagctt ctctgaccgt gaactacaga tacacctatg agggatctca catcaaggga
     3661 gaggctcagg tgaaaggaac tggattccct gctgatggac ctgtgatgac caactctctg
     3721 accgctgctg actggtgcag aagcaagaag acctacccaa atgacaagac catcatcagc
     3781 accttcaagt ggtcttacac cactggaaat ggcaagagat acagaagcac tgccagaacc
     3841 acctacacct tcgccaagcc aatggctgcc aactacctga agaaccagcc aatgtatgtg
     3901 ttcagaaaga ccgagctgaa acacagcaag accgagctga acttcaagga gtggcagaag
     3961 gctttcactg atgtgatggg aatggatgag ctgtacaagt aactcgagcc tctagtaatt
     4021 aagtctcagc caccgttaac tgaacatgtc aaaacctgtg gagactgttg agatttgatg
     4081 ttctgaaaag ataaagccta taaataaaat gttgcccaaa tttcctgcct gatgtttttc
     4141 tttgtctttg ctacatggct ttgctgctcg gatcggctca ctctgtgtat gccacgttca
     4201 ctttgtactc tccttctcac ggtaggttta ttatttttag atgtgcagtt agtttctgtg
     4261 aaataacaca ccacacactg atattgtctg tgcattgact tggtgagtgc acattgtttt
     4321 tgatcttgac atatttatat ttgattgatc aggtgaactg tgtgaatcta aagtgctcca
     4381 tacagatgtt ctgcattgaa aatattctca ttttattagt ggaagtgagt gtatgccagc
     4441 ggccgcggta ccctgcagtc tagacatttc agttgacgaa gacaaacaaa gttctgttgt
     4501 gactatgggg ggggggggcg cctggggatg gtctcgcccg gggagtaatt cagggtagaa
     4561 ccgccactgc ctttagcgac attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4621 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4681 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4741 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     4801 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4861 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4921 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4981 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5041 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5101 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5161 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5221 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5281 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5341 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5401 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5461 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5521 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5581 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5641 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5701 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5761 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5821 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5881 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5941 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6001 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6061 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6121 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6181 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6241 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6301 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6361 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6421 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6721 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6781 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6841 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6901 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6961 agtgagcgag gaagcggaag a
//
