LOCUS       sequence-120548-        9194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For trans-synaptic labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9194)
  AUTHORS   Feinberg EH, Vanhoven MK, Bendesky A, Wang G, Fetter RD, Shen K, 
            Bargmann CI
  TITLE     GFP Reconstitution Across Synaptic Partners (GRASP) defines cell 
            contacts and synapses in living nervous systems.
  JOURNAL   Neuron. 2008 Feb 7. 57(3):353-63.
  PUBMED    18255029
REFERENCE   2  (bases 1 to 9194)
  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: "Neuron. 
            2008 Feb 7. 57(3):353-63."
COMMENT     Sequence Label: Pflp-18::NGL-1::spGFP11
FEATURES             Location/Qualifiers
     source          1..9194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3066..3077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3282..3329
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     regulatory      5510..5519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6763..6785)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6885..6904
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6986..7004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7072..7176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7177..7245,7246..8037)
                     /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(7395..7414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8208..8796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8697..8716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8950..8967
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9084..9105
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9120..9125,9126..9143,9144..9150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9158..9174
                     /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     9163..9185
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgca aatctgtcac atactgctcg aatcggaggt
       61 aggtttgaaa aacatgcaat tcatttccaa aatatgacga attttgaaaa aaaaaaatag
      121 aaaagaaact caattaaatt accttatttt ttctagtacg tcaattttga cttgcactta
      181 aaatctgcat tttaaaaatg aaaattcagt gcagctgcag tgcagtgcgg ctattatttt
      241 cggcattttt gcaaaactta cacaccaccg ttataagaaa atatttgtct ttttttttaa
      301 atttttattc agtcgttatt cgaaaaagac ttacagtacc ttgctgaatt ccttccgcat
      361 tgcgctaaaa tttaccgatt gcaaaataat cttacatgct atagtaataa aaaatttaca
      421 tacttgccat ttcatttcat ttaattattt catccaattt atcttgatca gtattactct
      481 ttgttaaaaa ttccactgtt caatttttac atttcagttg ttttaatatc gaagtgcaca
      541 agaagcgcga acataatgga caccatcgct caaaaattat gcttctagtt gtagttcctc
      601 gagcattttt gatctcaaaa gacatttcca agagcaataa ttgccatttt tgagctacaa
      661 gtaccatatc ctgctcataa ttcatgttcc accactccct aaagtttttt ctttggaatt
      721 ccccagactt tgcgcctaaa attctcgtta ttctctccaa ttcaattggc tcaattcaac
      781 tgactagatt tcagtgatgt aggtagcgcg gaactcagta ttaagagagg actcaactca
      841 gtttctaatc caatttcaac aaattctaag acaccgaaac gctgcaggtc tcgtagagcc
      901 tacgtgttga caggttcttc cttttcaatc tatttccgtc gatattccgg cgtcatttag
      961 ttgaaaatga gtgaaaaaat gcgatctgac gcgaattcac gtgaactcga aaattcgaga
     1021 atgtaatcat taagtttcca aacaccaagt tctctttcga aaccaagtca atagaaaaat
     1081 gagacaccca cgagaagaag gttggtccga aaaaagaaac taaagaagtt atttatttct
     1141 ctttttgatg tttgttgagc agaatatgat ttagttaagc acatctgatt tcaacgccga
     1201 tgaatgaggt cgtaggcgtc acgttcaatt tgatctgatt tgttcagtgc ttacgagtag
     1261 aaattctaaa tatgatgtta ctttttggtt ctgtaagatg gagctgggta tacatgtggt
     1321 gcacggcaat cgctatttgg caatttcagt catttcggca aattcggcaa ttttggcgaa
     1381 ttctcatatt ttggaaattt tggcaaatgc ggaaaatttc aaaaacaagc aattaccaaa
     1441 tttgccaaac gccgaaaaat ttaaaaaaaa accgacattt gctggtattt ttttttaatc
     1501 aggatgctaa ttatgtaagg ttaatctata aattcttgaa acacacttaa ctttgaaatt
     1561 gaaatcactg acatttcgcg cattcgccga taaaatctct aaataaaata atgtaaactg
     1621 tgtccagttt tggtacttgt ttagtatacc tggttcaaaa tattttccaa tgcgttttcc
     1681 atttatttat tttagaaatg atcatttaat taaaacgtaa aattcgcatt ttcaagctta
     1741 gaaacattaa tcctaagtcc aatatggaca aaaagagtag tgtattagta ccaagtaaga
     1801 accctaaaaa aaaaatattc taattaaatt tttggcaaaa tgccacattt ttcttttttt
     1861 tctcaacggg gaaaagttgt gtttcttccc tttttattaa attttttgtt ttccttaggc
     1921 taagtacaac ttgaaatgga acagaaatta cataaattta ttgctttcac attttgctct
     1981 tcttcactca aatcttggac tcttgctttt cacaaaaaaa accttataaa caaaaataag
     2041 acatctgaaa aaatgatgaa cttcctgtca gtttgttcca gtatccttct taaatttcct
     2101 tgttttccac atcacaaaaa tcctacatca cgcttcgact catttcctga cagtcatcac
     2161 atcaccaaat gaatttctat ttttacaata tagttgtagt atttttgtcg aacatatatt
     2221 gttctaacta acttcttaaa aaattgtgct cttgactgaa cacaaatttc tgcgtcgttt
     2281 cacgtttact tttctcaatt ttttaaaacc aaaatttaac tgcaaacttc tgcattgttc
     2341 taaaaactca caggttatct caaaaagata aatttcaaag tgagattttt ttaaaaaaat
     2401 tccggactgt tttttgattt tgatgttttt aatgaatttt taaaattctt tattagtgcc
     2461 tgaaattaac aataatttta aagtttgtca cactgcgaat aaattaatct cacatttcgt
     2521 atgtttagaa agatgcatga aactccgtcg ttcttatgaa gttaattaag acatggctaa
     2581 ttgaaagccc acatggtcat gaaaaagtaa aacttacaac tttttactga atattaacca
     2641 gtctctataa atatcccagc ttagccggaa tagggtcacg aaaacatgaa aataaagctt
     2701 gaacctattg ttatgatttt caatgccacc ttttgcagtt taatatacac tattactact
     2761 tctcagttca agttcacttc tgagacggaa cctttatttt ctaactttct ttaatcttac
     2821 tttttatgag ccgaatgttc tattttgtaa aatgatcaga tacaagccca aagctaattt
     2881 tgccaataag tgttagtaga tcaattgaat tacaatataa tgacctgggc ggaatagatt
     2941 tttgcactga tttttttcca agcttactct atgaatattc atagatcatc ttcaaaggta
     3001 gcgtgtgttg tagttgtgtt ttggcgggtt cgtgttattt gactcagctc aagtagatgt
     3061 tgtagtcttc cctcaatcaa cgcgtgggct tgaccctctt taaattgagt ggaaaccagc
     3121 caatctattt gaaatctacc cccgacccgg gattggccaa aggacccaaa ggtatgtttc
     3181 gaatgatact aacataacat agaacatttt caggaggacc cttggctagc atggaacgca
     3241 tttatcttct tctccttctt tttctgccca ggatacgatc ccgcgatcac atggtcctgc
     3301 atgagtatgt gaacgccgcc gggatcactg agctcggatc tggatctgga tctggatctg
     3361 gatctggatc ttatgatgtt cgatcagtca ccacatcctg gggcatggtg agaggtgaag
     3421 tggttagccc cgaaggagat gatctaccgc cagtagcaca gtatctaggc attccgtatg
     3481 gcgtggctcc tactggacag tatcgtttca acatggccat atctgccgcc aaatggacac
     3541 atatgccaaa ggacgcgcgc aaagtttctc cagtctgcat tcaaactgac atgccggaat
     3601 taagtgaaac aaaagcattc aaacatactt ctgcacagcg ttttgacttt aatcatcgac
     3661 tgcttccgca tctcaagaaa caatcggaag actgtctcta catgaatatc tacgttccgg
     3721 agagattaga aatctccagg gacagctact taccggtaat ggtaattgta cacggagaag
     3781 agtatggatg gggtacgggc aacgcattta atggtaccac gttggctgct tacggtcata
     3841 ttattgtagt cacattgaac tatagactgg gtgttttcgg gtttttggga cgatgtgaat
     3901 catcgtcttg ctctggtaat agtggaattt cggatctggt atcggctttg acaatgctca
     3961 atgttatttt gccttcattt ggtggagact ctaaatcggt cactcttgct ggttggggtt
     4021 ctggcgcttc tctggtttct cttcttatgg cttccccatt aacacaacct ggcagacgtc
     4081 tattccgacg agcaattctt ctggacggaa gcgctttatc gccctgggca atctctcaga
     4141 atcctcaaca gtactttatg cagcttgccg aggaacttgc gtgtgctcca aaaaacagaa
     4201 catcatcttt caacgacaat gtggacacca ttgtccgatg tatgcaagta cactcatcgg
     4261 aaaacatcac aaaagcggtt ctcaaaatcg atgttcccac gtttttgtcg ggattcgcgc
     4321 cgattgttga tggacagctc ataccaaata aaccgcaagt atcattctcc acccaatacg
     4381 gatcactctt ccgagaaatt gatttgctag taggaatcag ctcgaatcca tcccaccaca
     4441 tgatttccaa tgaggatctg aaagtaggca tctccaaaga aaagcggatg agaatctttc
     4501 ggtcgttggt acgaaatttg tatgatttcc acagggaaga aattcttgcg tcaatcatca
     4561 atgagtatac agattgggaa aatcccaggg accacccgaa atcaattcga aatggagttc
     4621 tgaatgctct tagcgacgta ctttacaccg cacctctcat tgaaacattg cgaagtcaca
     4681 gcgctgatga agttcgcaaa gaggcaaata cgttcatgtt tgcatttgcc catgagaccc
     4741 ggtcttggag ccaagagcaa ccaaacagtg gaataagggg ctctctatct ggagacattg
     4801 taccatacat tttcggatac ccattagctc aaggggatag tgaggagagg ttgtattcag
     4861 gatttaacac tgacgataaa ggcatttcaa aagttatgat gcattatgta tcaaactttg
     4921 taaaatcggg agatccatca aagccaaatc caatgtccaa aaacttccca atgggggacg
     4981 tgtttcattc aacagcctgg ccacaatttg accaaccaaa tagagaagct tatttggaaa
     5041 ttactgatcg ccctcgagtc aaaaactact atcggaatgc tcaagtagga ttttggaata
     5101 actttatccc gcaattgcac aagaatggaa aggaaacaga gcctgttgga gaagagcatc
     5161 atttgctgtc agatcatttc cgaaaagact cttactttgg aaaaacccgt catttttctt
     5221 catatgcgaa tcttccgttc ccgccaccac caatgccccc atcaccacca ccagagctta
     5281 caacaaagcc caaaccaagt gaatcgccca caactctcca aacaacaact gaaagtgaaa
     5341 aggccgccgc tggaagtttt actggaaaag ctctcggtgg cgttattttc atcggttgtg
     5401 gattcctcat tatgaacgtt tgcctattaa ttgctgttcg tagagaatgg ggaaagaagc
     5461 ggcggaacga gaagaagttt caactgcagt atcagactta taactccaac catggcggcg
     5521 gcgcggaaca atacaacagc ttaaactcgc cgttatccct catgccacca cctccgccac
     5581 cgctcaatgg tgttcacgac gacattttcg atcatcgggt gccgcatctt cggaacggat
     5641 ccacggttca cggcacgctt ccgaggcatt catttcagga acaggaaccc ttactatccg
     5701 catcgcacaa gaactcaact tcgatgcgac ccgcgggaat atcaccaacg tgtccacgtc
     5761 acggacgtgc cgcgcttgcg cttcaaaata gccgtttgac tgctgctcaa gcaccgacat
     5821 tggaagagat acaggtctaa gaattccaac tgagcgccgg tcgctaccat taccaacttg
     5881 tctggtgtca aaaataatag gggccgctgt catcagagta agtttaaact gagttctact
     5941 aactaacgag taatatttaa attttcagca tctcgcgccc gtgcctctga cttctaagtc
     6001 caattactct tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg
     6061 ttattatcaa aaaaacttct tcttaatttc tttgtttttt agcttctttt aagtcacctc
     6121 taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa
     6181 attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg
     6241 tacacttctt atgttttttt tacttctgat aaattttttt tgaaacatca tagaaaaaac
     6301 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     6361 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     6421 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     6481 tgggggtttc ccctattgtt tgtcaagagt ttcgaggacg gcgtttttct tgctaaaatc
     6541 acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag
     6601 tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct
     6661 taaatgacag aatacattcc caatatacca aacataactg tttcctacta gtcggccgta
     6721 cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6781 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6841 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6901 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6961 ccgcatcagg cggccttaat taagggcctc gtgatacgcc tatttttata ggttaatgtc
     7021 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7141 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7201 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7261 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7321 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7381 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7441 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7501 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7561 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7621 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7681 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7741 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7801 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7861 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7921 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7981 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8041 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8101 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8161 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8221 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8281 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8341 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8401 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8461 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8521 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8581 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8641 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8701 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8761 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8821 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8881 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8941 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     9001 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     9061 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9121 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9181 acaggaaaca gctn
//
