LOCUS       sequence-479866-        6063 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  Expresses cytoplasmic fluorogen activating protein dL5** fused to 
            mCer under UAS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 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:dL5-mCer3
FEATURES             Location/Qualifiers
     source          1..6063
                     /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"
     CDS             1588..2307
                     /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"
     CDS             join(2365..2367,2368..2370,2371..3084)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2410..2431)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2671..2690)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3018..3039
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3667..3684)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3667..3683)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3676..3698)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3892..3911)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4011..4033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4071..4089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4157..4261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4262..4330,4331..5122)
                     /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(4480..4499)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5293..5881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5782..5801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6035..6052
                     /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 accatggaat tcggcctttc tggcctgcag gccgtcgtta cccaagaacc tagtgttacc
     1621 gttagcccag gtggtactgt tatacttact tgtggaagtg gtacgggtgc cgtcacatct
     1681 ggtcattatg caaattggtt tcaacaaaaa ccaggacaag ctccaagagc tttgattttt
     1741 gatactgata agaagtattc ttggacccca ggtagatttt ctggatcttt gctgggagca
     1801 aaggcagctt tgacaatatc agatgctcag cctgaggacg aagccgagta ttactgttct
     1861 cttagcgacg tggatggcta cttgtttggc ggtggaacac aactgacggt tctgtccggt
     1921 ggtggcggct ctggtggcgg tggcagcggc ggtggtggtt ccggaggcgg cggttctcag
     1981 gctgtggtga ctcaggagcc gtcagtgact gtgtccccag gagggacagt cattctcact
     2041 tgtggctccg gcactggagc tgtcaccagt ggtcattatg ccaactggtt ccagcagaag
     2101 cctggccaag cccccagggc acttatattt gacaccgaca agaagtattc ctggacccct
     2161 ggccgattct caggctccct ccttggggcc aaggctgccc tgaccatctc ggatgcgcag
     2221 cctgaagatg aggctgagta ttactgttcg ctctccgacg ttgacggtta tctgttcgga
     2281 ggaggcaccc agctgaccgt cctctccgcg gccggtctgg cccatatggg aggcggtggg
     2341 tctggtggcg gtggcagcac cggtatggtg agcaagggcg aggagctgtt caccggggtg
     2401 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2461 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2521 aagctgcccg tgccctggcc caccctcgtg accaccctga gctggggcgt gcagtgcttc
     2581 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2641 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2701 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2761 gaggacggca acatcctggg gcacaagctg gagtacaacg ccatccacgg caacgtctat
     2821 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcggcctcaa ctgcaacatc
     2881 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2941 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc
     3001 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3061 ggcatggacg agctgtacaa gtaactcgag cctctagtaa ttaagtctca gccaccgtta
     3121 actgaacatg tcaaaacctg tggagactgt tgagatttga tgttctgaaa agataaagcc
     3181 tataaataaa atgttgccca aatttcctgc ctgatgtttt tctttgtctt tgctacatgg
     3241 ctttgctgct cggatcggct cactctgtgt atgccacgtt cactttgtac tctccttctc
     3301 acggtaggtt tattattttt agatgtgcag ttagtttctg tgaaataaca caccacacac
     3361 tgatattgtc tgtgcattga cttggtgagt gcacattgtt tttgatcttg acatatttat
     3421 atttgattga tcaggtgaac tgtgtgaatc taaagtgctc catacagatg ttctgcattg
     3481 aaaatattct cattttatta gtggaagtga gtgtatgcca gcggccgcgg taccctgcag
     3541 tctagacatt tcagttgacg aagacaaaca aagttctgtt gtgactatgg gggggggggg
     3601 cgcctgggga tggtctcgcc cggggagtaa ttcagggtag aaccgccact gcctttagcg
     3661 acattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3721 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3781 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     3841 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     3901 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3961 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4021 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4081 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4141 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4201 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4261 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4321 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4381 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4441 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4501 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4561 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4621 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4681 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4741 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4801 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4861 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4921 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4981 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5041 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5101 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5161 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5221 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5281 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5341 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5401 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5461 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5521 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5581 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5641 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5701 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5761 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5821 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5881 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5941 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6001 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6061 aga
//
