LOCUS       sequence-120545-        9503 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 9503)
  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 9503)
  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: Punc-4::NGL-1::spGFP1-10
FEATURES             Location/Qualifiers
     source          1..9503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3039..3060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5819..5828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(7072..7094)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7194..7213
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7295..7313)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7381..7485
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7486..7554,7555..8346)
                     /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(7704..7723)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8517..9105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9006..9025
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9259..9276
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9393..9414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9429..9434,9435..9452,9453..9459)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9467..9483
                     /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     9472..9494
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgca gctctgagac catgttgatg ggaaattata
       61 tttgaaaact tttgaataaa gttgtataat tgtttttatt ctttaaccta tttatgaatg
      121 ctgtcccgga actgggatat aatttctaac ttgacacaat tattccacct ctgtcttcaa
      181 ggcgacctac acctacctcg cgtctaggtc caacagaaac actgttgaat gaaatcctcg
      241 aattgcaaca atataggatg ctcaggagac tataattcaa aaaacccaac ccaaattagc
      301 ataagcttcc ccaaattgga acagtgaaat actcaatcaa tcactctggc ggttttctcc
      361 acaaagaaca cgaaattgga aaaaggaaaa gattgatttg tttgaaggcg tgaggaaatc
      421 gaaaaaaaat tgtttcaaaa caaatgaaat attttaacga ataaacccag gaatgccact
      481 ttttggtgcc cgcctacaaa aaagtcaact gttcttaaac tcctacatag ttgccctctc
      541 taacgaatgt tgcttttttt aaattttcat tgaatttgat tatcgtctgt atgcgcctac
      601 tctatcagtg gaaaacatgc ccatttccca caaatttact cgtttggagc tcaaaatgtt
      661 aaatggggca tcagggaatt ggcatctcac agaaagtggc ggatgagtta tatgtaataa
      721 taatggtggc tgaagtggaa accgcaaaaa aaggatgtgt atttgagatt ttgcaacgga
      781 atttgaatca gatacacaaa atgagagaaa tttatagatt ctttaaaatc cacctgaagt
      841 cggcaaattt ttgaaaggtc tgttcgttca gtttttcata agttttatga attttgattt
      901 actactgaag ttcgagaaaa gccctttaac cataccaata agaaattatg ttaactttac
      961 aagaatatta acaaaaaatt attaaaaaac tgtgttcccc tggcattatg atatttccac
     1021 cgtcagttct ctattaaaac ttcaactcta cacagtttca actctcaaat gagctgtcca
     1081 tcttttgcct tcaggaaatt atgctaatct gctgatttcg cgcactttca ccacactatt
     1141 tccaaataga aggaggggtg gtgcaacaca cgagacactg caaaaaggtt cagtgaagat
     1201 ttatctactt gtgagatgag agcaggaggg gtgggagaag tgtgatgatt gacgggatga
     1261 caagaacagt gatgtggtgt cgaaatggaa gatgcgctaa tttgtggagt tttatgaaat
     1321 ttgaataatt acatttataa catctactag ttaggcgtct gtgtaggtat tgcctgcctg
     1381 cctacaaggc atatgtacga ggcaggttaa acctcctaga aagctacggt actttcaaac
     1441 cacataacat ttatactggt gtcttttcgc gtcttcttct tgaacatgta gaccctcaaa
     1501 aagctaagtt ttacactgtg tagtttgctt tgaaatctac aaaaattgga ggcacgcctt
     1561 ctggactttg ccgacacgcc tactttctac ctaccaactg cctacttcca cagtctagcg
     1621 tgaactccaa taccatttta aaaattgttt tatctcaaat tttcaatttt taaatccaat
     1681 ttcccagttc taaaaaaaaa attttttttt taatttaaaa actctcatcc tttttaatgc
     1741 tcgttcccat tcggcaagac gtgtaccaaa cacaaaacaa attgactggt gatccatctc
     1801 gaatttctcg tttcaaatta gtcgttgcta tatatattgt cttatgtgtg gaaaacgaat
     1861 aagaaaaatt agagaaagat gagagagcag gggcagaatg aaaatttgca tggaaattga
     1921 agaagatcac tgctttttcc gatgaatgat gactaaatag aatgagttga gaggaaaata
     1981 atcataattc taattttaat tagaaatatg atatggaaag taagcaaaac cacaaggctt
     2041 ttgctctgaa cttatttccg aaaaaaaaag tgaaactgaa agttagaatt ttcaaaataa
     2101 atattgcgag agtcaacctt cagaaaaatt ggcgagaaat caaatttgaa ttctctgaaa
     2161 atttagttaa aatacatatg ttatccttct ttgttttttg gccttagcct gccttcaagg
     2221 cgacctgagc ctatgcgggt gcctaatacc gaaacacaca taattttttt ttttaatctt
     2281 taaatactgt aacgagtcgc gagatccagc gagacccata gaatatgttg agcgctgcaa
     2341 acaatcgtat aactcttgtc caaatttttc cactctgtat ttttgcaagc agttcaacaa
     2401 atggtcatca tttttataac ccatatctat attcgaatct gacataacaa ataatactaa
     2461 gcacatcctc ccacatctct tttcattcaa gttgcgcaat tcccctaata actatggatg
     2521 gaggggggac cgagcccaaa gattattcaa aacgataacg actttttcat gcggccaacg
     2581 aaaaataaaa cttttctcat attctcctat ttcccatttc aagaattaca tccccatggg
     2641 acgttgaaat cacatcatat tagtatatgt taattgttct tttcactgga atatcgccac
     2701 gccccccttc cggtggctca ttggtttcga gaagaaggcg gagccgaagt gggttcttct
     2761 ttcttttttt tgtggtagaa gtctagcagg attaagctac tattccatta gaggatcttc
     2821 ttcttccaaa aagtgaaagg cgcgcctcta gaggatcccc gggattggcc aaaggaccca
     2881 aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga cccttggcta
     2941 gcatggaacg catttatctt cttctccttc tttttctgcc caggatacga tccatggtga
     3001 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3061 taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc attggcaagc
     3121 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3181 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     3241 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg
     3301 acgacggcaa atacaagacc cgcgccgtgg tgaagttcga gggcgacacc ctggtgaacc
     3361 gcatcgagct gaagggcacc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3421 agtacaactt taacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     3481 aggcgaactt caccgtgcgc cacaacgtgg aggacggcag cgtgcagctc gccgaccact
     3541 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3601 gcacccagac cgtgctgagc aaagacccca acgagaagga gctcggatct ggatctggat
     3661 ctggatctgg atctggatct tatgatgttc gatcagtcac cacatcctgg ggcatggtga
     3721 gaggtgaagt ggttagcccc gaaggagatg atctaccgcc agtagcacag tatctaggca
     3781 ttccgtatgg cgtggctcct actggacagt atcgtttcaa catggccata tctgccgcca
     3841 aatggacaca tatgccaaag gacgcgcgca aagtttctcc agtctgcatt caaactgaca
     3901 tgccggaatt aagtgaaaca aaagcattca aacatacttc tgcacagcgt tttgacttta
     3961 atcatcgact gcttccgcat ctcaagaaac aatcggaaga ctgtctctac atgaatatct
     4021 acgttccgga gagattagaa atctccaggg acagctactt accggtaatg gtaattgtac
     4081 acggagaaga gtatggatgg ggtacgggca acgcatttaa tggtaccacg ttggctgctt
     4141 acggtcatat tattgtagtc acattgaact atagactggg tgttttcggg tttttgggac
     4201 gatgtgaatc atcgtcttgc tctggtaata gtggaatttc ggatctggta tcggctttga
     4261 caatgctcaa tgttattttg ccttcatttg gtggagactc taaatcggtc actcttgctg
     4321 gttggggttc tggcgcttct ctggtttctc ttcttatggc ttccccatta acacaacctg
     4381 gcagacgtct attccgacga gcaattcttc tggacggaag cgctttatcg ccctgggcaa
     4441 tctctcagaa tcctcaacag tactttatgc agcttgccga ggaacttgcg tgtgctccaa
     4501 aaaacagaac atcatctttc aacgacaatg tggacaccat tgtccgatgt atgcaagtac
     4561 actcatcgga aaacatcaca aaagcggttc tcaaaatcga tgttcccacg tttttgtcgg
     4621 gattcgcgcc gattgttgat ggacagctca taccaaataa accgcaagta tcattctcca
     4681 cccaatacgg atcactcttc cgagaaattg atttgctagt aggaatcagc tcgaatccat
     4741 cccaccacat gatttccaat gaggatctga aagtaggcat ctccaaagaa aagcggatga
     4801 gaatctttcg gtcgttggta cgaaatttgt atgatttcca cagggaagaa attcttgcgt
     4861 caatcatcaa tgagtataca gattgggaaa atcccaggga ccacccgaaa tcaattcgaa
     4921 atggagttct gaatgctctt agcgacgtac tttacaccgc acctctcatt gaaacattgc
     4981 gaagtcacag cgctgatgaa gttcgcaaag aggcaaatac gttcatgttt gcatttgccc
     5041 atgagacccg gtcttggagc caagagcaac caaacagtgg aataaggggc tctctatctg
     5101 gagacattgt accatacatt ttcggatacc cattagctca aggggatagt gaggagaggt
     5161 tgtattcagg atttaacact gacgataaag gcatttcaaa agttatgatg cattatgtat
     5221 caaactttgt aaaatcggga gatccatcaa agccaaatcc aatgtccaaa aacttcccaa
     5281 tgggggacgt gtttcattca acagcctggc cacaatttga ccaaccaaat agagaagctt
     5341 atttggaaat tactgatcgc cctcgagtca aaaactacta tcggaatgct caagtaggat
     5401 tttggaataa ctttatcccg caattgcaca agaatggaaa ggaaacagag cctgttggag
     5461 aagagcatca tttgctgtca gatcatttcc gaaaagactc ttactttgga aaaacccgtc
     5521 atttttcttc atatgcgaat cttccgttcc cgccaccacc aatgccccca tcaccaccac
     5581 cagagcttac aacaaagccc aaaccaagtg aatcgcccac aactctccaa acaacaactg
     5641 aaagtgaaaa ggccgccgct ggaagtttta ctggaaaagc tctcggtggc gttattttca
     5701 tcggttgtgg attcctcatt atgaacgttt gcctattaat tgctgttcgt agagaatggg
     5761 gaaagaagcg gcggaacgag aagaagtttc aactgcagta tcagacttat aactccaacc
     5821 atggcggcgg cgcggaacaa tacaacagct taaactcgcc gttatccctc atgccaccac
     5881 ctccgccacc gctcaatggt gttcacgacg acattttcga tcatcgggtg ccgcatcttc
     5941 ggaacggatc cacggttcac ggcacgcttc cgaggcattc atttcaggaa caggaaccct
     6001 tactatccgc atcgcacaag aactcaactt cgatgcgacc cgcgggaata tcaccaacgt
     6061 gtccacgtca cggacgtgcc gcgcttgcgc ttcaaaatag ccgtttgact gctgctcaag
     6121 caccgacatt ggaagagata caggtctaag aattccaact gagcgccggt cgctaccatt
     6181 accaacttgt ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa gtttaaactg
     6241 agttctacta actaacgagt aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac
     6301 ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc
     6361 ctatttttgt tattatcaaa aaaacttctt cttaatttct ttgtttttta gcttctttta
     6421 agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag
     6481 tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa
     6541 tgttctgtgt acacttctta tgtttttttt acttctgata aatttttttt gaaacatcat
     6601 agaaaaaacc gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt
     6661 tttatttctt cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt
     6721 tttggagtat ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct
     6781 tttgcttttt gggggtttcc cctattgttt gtcaagagtt tcgaggacgg cgtttttctt
     6841 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     6901 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     6961 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     7021 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7081 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7141 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7201 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7261 gagaaaatac cgcatcaggc ggccttaatt aagggcctcg tgatacgcct atttttatag
     7321 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     7381 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     7441 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     7501 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     7561 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     7621 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     7681 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     7741 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     7801 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     7861 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7921 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7981 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     8041 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     8101 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     8161 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     8221 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     8281 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     8341 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     8401 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     8461 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     8521 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     8581 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     8641 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     8701 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8761 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8821 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8881 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     8941 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     9001 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     9061 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     9121 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     9181 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     9241 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     9301 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     9361 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     9421 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     9481 caatttcaca caggaaacag ctn
//
