LOCUS       sequence-401573-        8515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing an antigen targeting Neuroligin-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8515)
  TITLE     Trimmer lab antigen plasmids
REFERENCE   2  (bases 1 to 8515)
  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"
COMMENT     Sequence Label: Neuroligin-3 GFP/pCMV
FEATURES             Location/Qualifiers
     source          1..8515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(75..95)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        80..409
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      260..395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     322..341
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             476..1270
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(530..549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1140..1159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1444..1565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1481..1500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1535..1554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1614..1630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1614..1630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1627..1649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1638..1654
                     /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)."
     promoter        complement(join(1662..1668,1669..1686,1687..1692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1707..1728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1845..1862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2016..2604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2096..2115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2775..3566,3567..3635))
                     /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     3398..3417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3636..3740)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3815..3834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4006..4385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4386..4589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4540..4560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4634..4653
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4634..4652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4689..4733
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             4734..4757
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(6996..6998,6999..7001,7002..7712)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7041..7062)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7302..7321)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7649..7670
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(7839..7857)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7840..7857)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7877..7894)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7883..8107
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8153..8515,1..66)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8240..8259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8450..8471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
       61 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
      121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      181 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
      361 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
      421 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
      481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
      901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
      961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     1021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     1081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     1141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     1201 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1261 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     1321 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1381 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1441 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1501 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1561 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     1621 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1681 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2761 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2821 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2881 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2941 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3001 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3061 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3121 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3181 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3241 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3301 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3361 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3421 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3481 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3541 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3601 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3661 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3721 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     3781 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3841 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     3901 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     3961 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     4021 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4081 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4141 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4201 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4261 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4321 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4381 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4441 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4501 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4561 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     4621 ggcttatcga aattaatacg actcactata gggagaccca agcttggtac cgagctcgga
     4681 tcctcaccat gtctgcactt ctgatcctag ctcttgttgg agctgcagtt gctgactaca
     4741 aagacgatga cgacaagctt gcggccgcca gtacccaggc cccggcaccc acagtcaata
     4801 ctcactttgg gaaactaagg ggtgccagag taccattgcc cagtgaaatc ctgggtcctg
     4861 tggaccaata cctgggggta ccctacgcag ctcccccgat cggcgagaaa cgtttcctgc
     4921 cccctgaacc acccccatcc tggtcgggca tccggaacgc cacacacttt cccccagtgt
     4981 gcccccagaa catccacaca gctgtgcccg aagtcatgct gccagtctgg ttcactgcca
     5041 acttggatat cgtcgccact tatatccagg agcccaacga agattgcctc tatctgaatg
     5101 tgtatgtgcc cacggaagat gacatccgag acagtggtgc taaacctgtc atggtctaca
     5161 tccacggagg ctcttacatg gaaggaacag gcaacatgat tgacggcagc gttcttgcaa
     5221 gttatggcaa cgtcatcgtc atcacactca actaccgggt cggggtgcta ggtttcctga
     5281 gcactgggga tcaggctgcc aagggcaact atgggctcct tgatcaaatc caggcccttc
     5341 gctgggtgag tgagaacatt gccttctttg gaggagatcc ccgtagaatt actgtctttg
     5401 gctctggcat tggtgcatcc tgtgtcagtc ttctcacact gtctcatcac tctgaggggc
     5461 ttttccagag ggccatcatc caaagtggct cagcgctatc tagctgggct gtgaactacc
     5521 aaccagtgaa gtacaccagc ttgctcgcag acaaagtggg ctgtaacgtc ctggacactg
     5581 tggatatggt ggattgtctt cgacaaaaga gtgccaagga gctggtagaa caggacattc
     5641 agccagcccg ctaccatgtg gcttttggcc ctgtgattga tggtgatgtc atttctgatg
     5701 accctgagat ccttatggaa cagggagagt tcctcaacta tgatatcatg ctaggcgtca
     5761 accagggtga gggtctcaag tttgtggaag gggtggtgga ccccgaggat ggtgtctctg
     5821 gcactgactt tgactactct gtctccaatt ttgtggacaa tctatatggc tatcccgagg
     5881 gtaaggacac cctgcgggag actatcaagt tcatgtatac agactgggca gaccgagaca
     5941 accctgagac ccgccgtaaa acactggtgg cactcttcac tgaccaccag tgggtggagc
     6001 cttcggtggt gacagctgat ctgcatgccc gctatggctc acccacttac ttctacgcct
     6061 tctaccatca ctgccagagc ctcatgaagc ctgcatggtc agatgcagca cacggggatg
     6121 aagtacccta tgtttttggt gtccctatgg taggtcccac tgaccttttc ccctgcaact
     6181 tctccaagaa tgatgttatg ctcagtgctg ttgttatgac ctattggacc aactttgcca
     6241 agaccgggga tcccaacaag ccggtacccc aggataccaa gttcattcac accaaggcca
     6301 accgctttga ggaagtagcc tggtccaaat acaatccccg ggaccagctc taccttcaca
     6361 tcgggctgaa accaagggtt cgtgatcatt accgagccac aaaggtagcc ttttggaagc
     6421 acctggtacc ccatctgtac aacctgcatg acatgttcca ctatacatcc acgaccacga
     6481 aagtgccgcc gccggacacc acccatagct cccacatcac ccgtaggccc aacggcaaga
     6541 cctggagcac caagaggcca gccatctcac ctgcctacag caatgagaat gctcctgggt
     6601 cctggaatgg ggaccaggat gcagggccac tcctggtcga gaaccctcga gactactcca
     6661 ctgaattaag tgtcaccatc gctgttgggg cctctctcct gtttctcaat gtgttggcct
     6721 ttgctgctct ctattaccgt aaggacaaac ggcgccagga gcccctgagg cagcctagcc
     6781 cccaaagggg aactggtgcc ccagaattgg gaactgctcc agaggaggag ctggcagcat
     6841 tacagttggg tcccactcac catgagtgtg aggccggccc cccacacgac acactgcgcc
     6901 tcacagcatt gcccgactac accctgaccc tgcggcgctc ccccgatgac atcccactca
     6961 tgactcccaa caccatcact atgattccga attccatggt gagcaagggc gaggagctgt
     7021 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     7081 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     7141 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc
     7201 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     7261 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     7321 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     7381 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     7441 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     7501 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     7561 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctta
     7621 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     7681 ggatcactct cggcatggac gagctgtaca agaattccct ggttgggttg cagaccttgc
     7741 acccctataa cacctttgcc gccggattca acagcaccgg gctgccccac tcacactcca
     7801 ctacccgtgt atagctcgtc gactctagag ggccctattc tatagtgtca cctaaatgct
     7861 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     7921 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     7981 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     8041 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     8101 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     8161 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8221 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8281 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8341 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8401 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     8461 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataag
//
