LOCUS       sequence-120558-        8254 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 8254)
  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 8254)
  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::spGFP1-10
FEATURES             Location/Qualifiers
     source          1..8254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5823..5845)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5945..5964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6046..6064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6132..6236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6237..6305,6306..7097)
                     /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(6455..6474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7268..7856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7757..7776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8010..8027
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8144..8165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8180..8185,8186..8203,8204..8210)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8218..8234
                     /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     8223..8245
                     /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 ctgctagcat gtccaaagga gaagaactgt ttaccggtgt tgtgccaatt
     3181 ttggttgaac tcgatggtga tgtcaacgga cataagttct cagtgagagg cgaaggagaa
     3241 ggtgacgcca ccattggaaa attgactctt aaattcatct gtgtaagttt aaacaataac
     3301 ctactaacta acgtagataa tttaaatttt cagactactg gtaaacttcc tgtaccatgg
     3361 ccgactctcg taacaacgct tacgtacgga gttcagtgct tttcgagata cccagaccat
     3421 atgaaaagac atgacttttt taagtcggct atgcctgaag gttacgtgca agaaagaaca
     3481 attgtaagtt taaacttgat agtactaact aacatgtttc atttaaattt tcagtcgttc
     3541 aaagatgatg gaaaatataa aactagagca gttgttaaat ttgaaggaga tactttggtt
     3601 aaccgcattg aactgaaagg aacagatttt aaagaagatg gtaatattct tggacacaaa
     3661 ctcgaataca attttaatag tcatgtaagt ttaaactgag ttctactaac taacgagtaa
     3721 tatttaaatt ttcagaacgt atacatcact gctgataagc aaaagaacgg aattaaagcg
     3781 aatttcacag tacgccataa tgtagaagat ggcagtgttc aacttgccga ccattaccaa
     3841 caaaacaccc ctattggaga cggtccggta cttcttcctg ataatgtaag tttaaacagg
     3901 atcttactaa ctaacatgct aacactgaat tttcagcact acctctcaac acaaacagtc
     3961 ctgagcaaag atccaaatga aaaaggaaca gtcgacggag gaggaggatc aggaggagga
     4021 ggatctttcc aaaaagcatc atctattgtt tataagaagg aaggtgagta agtttaaaca
     4081 gatccatact aactaacttg ttctgacata attttcagac aagtcgagtt ttcattcccg
     4141 cttgccttta ctgttgagaa acttactgga tctggagaac tttggtggca agccgaaaga
     4201 gcttcttcat caaagtcatg gattactttt gatttgaaaa ataaagaggt aagtttaaac
     4261 agttgaatac taactaacgg agatctttga aattttcagg tgtcagtgaa aagagtgaca
     4321 caagatccga aattgcagat gggaaagaaa cttccacttc atttgacttt gccacaagcc
     4381 ttgccacagt atgcaggttc aggaaatttg acacttgctt tggaagccaa gtaagtttaa
     4441 acaataacct actaactaac gtagataatt taaattttca gaacaggaaa gcttcaccaa
     4501 gaggtcaatc ttgtggtgat gcgagctact caattgcaga agaatcttac atgcgaagtt
     4561 tggggaccaa catctccaaa attgatgttg tcattgaaac ttgaaaacaa gtaagtttaa
     4621 acttgatagt actaactaac atgtttcatt taaattttca gggaggcaaa agtgtctaag
     4681 cgtgagaaag ctgtttgggt tcttaaccca gaagcaggaa tgtggcagtg tttgttgtca
     4741 gattctggtc aagtgttgct tgaatcaaat attaaggtct tgccgacatg ggtaagttta
     4801 aactgagttc tactaactaa cgagtaatat ttaaattttc agtctacacc ggttcagcca
     4861 atggcactta ttgttcttgg tggtgtggca ggacttttgt tgttcatcgg attgggaatc
     4921 ttcttctgtg tccgatgccg acatcgacgt cgttaaggta ccgacgtcat gcgaagggcg
     4981 aattctgcag atatctgagc tccgcatcgg ccgctgtcat cagatcgcca tctcgcgccc
     5041 gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt
     5101 gctcccaccc cctatttttg ttattatcaa aaaaacttct tcttaatttc tttgtttttt
     5161 agcttctttt aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc
     5221 gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa
     5281 aatctacaca atgttctgtg tacacttctt atgttttttt tacttctgat aaattttttt
     5341 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     5401 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     5461 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     5521 attttcctgc ttttgctttt tgggggtttc ccctattgtt tgtcaagagt ttcgaggacg
     5581 gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa
     5641 ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag
     5701 tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca aacataactg
     5761 tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5821 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5881 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5941 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6001 agatgcgtaa ggagaaaata ccgcatcagg cggccttaat taagggcctc gtgatacgcc
     6061 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6121 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6181 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6241 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6301 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6361 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6421 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6481 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6541 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6601 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6661 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6721 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6781 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6841 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6901 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6961 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7021 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7081 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7141 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7201 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7261 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7321 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7381 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7441 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7501 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7561 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7621 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     7681 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7741 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7801 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7861 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7921 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7981 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8041 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8101 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8161 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     8221 tgagcggata acaatttcac acaggaaaca gctn
//
