LOCUS       sequence-120559-        7444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For membrane contact.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  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 7444)
  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: Prig-3::CD4::spGFP11
FEATURES             Location/Qualifiers
     source          1..7444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3139..3186
                     /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"
     primer_bind     complement(5013..5035)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5135..5154
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5236..5254)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5322..5426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5427..5495,5496..6287)
                     /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(5645..5664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6458..7046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6947..6966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7200..7217
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7334..7355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7370..7375,7376..7393,7394..7400)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7408..7424
                     /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     7413..7435
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gccgaaatgc
       61 gacccatcat attgttccta ttgtcatgta caagctgtcc ctgttttgct cgcttaatcc
      121 gtcaacccct tccccgttct aattagagat gagtgaatta caatagagtt gaaacggaaa
      181 tctgagtgtc tgtagctttc cgccgccgct tttgttacga gatggtagac gtccctttct
      241 tggtgtttat tgtctcacaa actgtataat aaagcaccgt tgtgtaaaag gcatttggta
      301 tcaggaagat aaaatgaata aaatgcaaat gcggaatcaa aaattgccaa ataattttga
      361 ccaataaaat atcaaaaata atagaacaga aataaacaaa taaaactacg gagaaaatgt
      421 gattattcca ttaaacaacg ttttaaacca actttattcg tcagtgggga aggttttgga
      481 caaatatcat agtaaaactt gttcgtttta attatacata atcaataatt ataggtgtta
      541 gaaccaatgg tcgaaattta aaaaaaaatc aacattttag aaaatttcgg caatttaccg
      601 aaatattgac tgacaatttt acatatataa gtgttttgaa atagaatccc cattgacgat
      661 gcgccacctt aaacagtaga actgttccaa tcagaaacta aataaatacg gttattatat
      721 atatatatat atatatatat atataacttg tttttattat gcatttctca aaactcacca
      781 aactcaatct ctaatttacg gttgttgagg aaaatatatt tatcacattt agttgagact
      841 ctaaaagaag tacaactttt ttcttgcaat tcctccacca atccaatttt tcaagagtca
      901 attgcttgga gtcctcgttt tctcgtaatc cgatattccc cctatttctg tgattttatt
      961 cagtttcgtc ctcttacacc aaaaatcttc atgacttttt tggttttctg cttaatcttt
     1021 tttttgtttt taaagtctac catccggcaa ttttttttgc aaatggttgc tcagaatttg
     1081 acctcagatt aaaattttca ggaacttcct ggtcccattt cgtcacattt aataacaaac
     1141 caaatccttc agcttgagtc gtccaagcca acgcaaccct aattcgattc cacctccaat
     1201 ttaccttttt gccgttttcg atcaaattgt gggggctctt tttatttgaa agtcgtaaaa
     1261 tggcgcgtcc ttgtcgaaaa ttgagatagc taatggttcc aaataaaatc aaatgcatga
     1321 acctagtagg aaaaatcgtt ttttttttca tcttccaaag aaaaagttta agataaaccc
     1381 accactccag caatccacct acgcgttttt caccggaaac tgtcatttgt ttttgatgta
     1441 tgacagtgcc agcactgaac taaattccca tttgttttgt tgtggtgtta tctaaccacg
     1501 tggctctttc tctaaatttt gcgtaggctg caagatccgc aatcccacca aaaaatggtt
     1561 tctaatgtac ttcatacata tcccacatga ctcagtgctc cgaaacgccc agcaaccccc
     1621 gcgtttgtct ttacaaaaat gaggcggaaa tgggaaattt tcctgctatt gctatggtaa
     1681 aatacttaag ctgtattgag atttgggaaa ttgaaagata agatgaaaaa tccactctgt
     1741 ataattaaaa aggtttattt tttttgtaat ttttgtaccg caaatttttt cattaactga
     1801 tttttcttta ttcattcgat ggcccagttt cagatcttca ttatcatgta gtttattcat
     1861 tcaaactatt tgtgacatca tgatggttgt ttaaaaataa tttcgaatga agaaagctta
     1921 aacctattat aattttcttt tctataaaaa atattgattt tgtaaagcgt ttggcagtga
     1981 cagtaccaaa aaatgtttta aaatgattta agaaaatttt ttattttatt tgtataatac
     2041 gcgtaaaatg agtatactcg tcatttgtca cgaatcgagt tctcagtttg ctgtgaatat
     2101 cttgtaatag aaatcgatct catagtgtca cttttgaggc ataacttttc aggttgttac
     2161 aagttcatta acttgtcaag actgggtcat gttttaaagt gtaactgtta cccatgtacc
     2221 ctaaactttg agacaaactc ctcaaaactt gtaaccagaa tcctatgtag atgactcaca
     2281 aaaccctgtt ttcaatttga acgaatttcg aaactggcca aaatgttccc ctcttgctca
     2341 cctaatgctt ttaccattta aaaaggatgg taacaacact ctttcgaaaa tgtaggcggt
     2401 ccattttcgg tttctcataa ccgtctggct ctatctccta gtgcgcgagg agagggcggg
     2461 cggagtagga atcgggggca gcgcactcat acactctgcc cttctctttt gcacacacag
     2521 aaaaagatag atacagacca acacttattg ttgtccagta agcttccaga aaactatttt
     2581 cagaattaca aactacattt ttctagaaca gtcaattgat tttttacaag attcgagtat
     2641 tctaacatta gtgaaagtat ttagccactg caattttcaa ctgtaggttt caacttccaa
     2701 aaaaaattcc tagcatttct aattggatga aaaaaggcta tctagaccct ccattattta
     2761 ctcataattt catctcgtcc tcatttttaa tcccctcatt gatcccatca caatccgcca
     2821 aaatgttttt taaacacttt tttcttcacc agccgtatgt ggtcaactgc ctaaaaattt
     2881 cacagctatt tgtcattcct tgcagaagaa cttcattctt ctttttcgaa aggcgcgcct
     2941 ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat
     3001 aacatagaac attttcagga ggacccttgg agggtaccgc cggcaatgcc accttcaaca
     3061 tcattgctgc tcctcgcagc acttcttcca ttcgctttac cagcaagcga ttggaagact
     3121 ggagaagtca ctgctagcag agatcatatg gtgttgcatg aatatgtcaa tgctgcaggt
     3181 attactggag gaggaggatc aggaggtgga ggatctttcc aaaaagcatc atctattgtt
     3241 tataagaagg aaggtgagta agtttaaaca gatccatact aactaacttg ttctgacata
     3301 attttcagac aagtcgagtt ttcattcccg cttgccttta ctgttgagaa acttactgga
     3361 tctggagaac tttggtggca agccgaaaga gcttcttcat caaagtcatg gattactttt
     3421 gatttgaaaa ataaagaggt aagtttaaac agttgaatac taactaacgg agatctttga
     3481 aattttcagg tgtcagtgaa aagagtgaca caagatccga aattgcagat gggaaagaaa
     3541 cttccacttc atttgacttt gccacaagcc ttgccacagt atgcaggttc aggaaatttg
     3601 acacttgctt tggaagccaa gtaagtttaa acaataacct actaactaac gtagataatt
     3661 taaattttca gaacaggaaa gcttcaccaa gaggtcaatc ttgtggtgat gcgagctact
     3721 caattgcaga agaatcttac atgcgaagtt tggggaccaa catctccaaa attgatgttg
     3781 tcattgaaac ttgaaaacaa gtaagtttaa acttgatagt actaactaac atgtttcatt
     3841 taaattttca gggaggcaaa agtgtctaag cgtgagaaag ctgtttgggt tcttaaccca
     3901 gaagcaggaa tgtggcagtg tttgttgtca gattctggtc aagtgttgct tgaatcaaat
     3961 attaaggtct tgccgacatg ggtaagttta aactgagttc tactaactaa cgagtaatat
     4021 ttaaattttc agtctacacc ggttcagcca atggcactta ttgttcttgg tggtgtggca
     4081 ggacttttgt tgttcatcgg attgggaatc ttcttctgtg tccgatgccg acatcgacgt
     4141 cgttaaggta ccgacgtcat gcgaagggcg aattctgcag atatctgagc tccgcatcgg
     4201 ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga cttctaagtc caattactct
     4261 tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa
     4321 aaaaacttct tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa
     4381 attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc
     4441 ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt
     4501 atgttttttt tacttctgat aaattttttt tgaaacatca tagaaaaaac cgcacacaaa
     4561 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct
     4621 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat
     4681 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tgggggtttc
     4741 ccctattgtt tgtcaagagt ttcgaggacg gcgtttttct tgctaaaatc acaagtattg
     4801 atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga
     4861 gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag
     4921 aatacattcc caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt
     4981 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5041 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5101 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5161 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5221 cggccttaat taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5281 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5341 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5401 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5461 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5521 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5581 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5641 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5701 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5761 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5821 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5881 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5941 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6001 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6061 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6121 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6181 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6241 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6301 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6361 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6421 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6481 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6541 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6601 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6661 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6721 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6781 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6841 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6901 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6961 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7021 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7081 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7141 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7201 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7261 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7321 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     7381 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     7441 gctn
//
