LOCUS       sequence-226833-        8457 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of PSD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8457)
  TITLE     Kahn expression plasmids
REFERENCE   2  (bases 1 to 8457)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: g15s pD47
FEATURES             Location/Qualifiers
     source          1..8457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        555..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     709..729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     803..822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        803..821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    854..878
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3210..3234)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3245..3247,3248..3250,3251..3964)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3278..3306)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3901..3921
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3986..4003)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3992..4216
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4262..4690
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4349..4368)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4559..4580
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4699..4719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4704..5033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4884..5019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5100..5894
                     /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(5154..5173)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5764..5783
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6070..6191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6107..6126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6161..6180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6240..6256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6240..6256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6253..6275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6264..6280
                     /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(6288..6294,6295..6312,6313..6318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6333..6354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6471..6488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6642..7230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6722..6741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7401..8192,8193..8261))
                     /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     8024..8043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8262..8366)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(8441..8457,1..3))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
       61 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      121 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      181 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      541 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      721 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      781 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      841 taagcttgat caaacaagtt tgtacaaaaa agcaggctta gaaggagata gaaccatgga
      901 ggaggacaag ctcttatctg cagtgcctga ggaaggcgat gccacccgtg accccggtcc
      961 agagcctgaa gaggagccag gggtccggaa tgggatggcc agtgagggcc tgaacagcag
     1021 cctctgcagc ccagggcacg agcgaagggg caccccagcg gacactgagg aacccacgaa
     1081 ggacccagat gtggccttcc atggcctcag ccttggcctc tctctcacca atggcctagc
     1141 cctggggcca gacttgaaca ttctggaaga ttcagcggag tccaggccct ggagggctgg
     1201 cgtgctggca gagggggaca atgcttccag gagcctctac ccagatgctg aggaccctca
     1261 gctggggttg gatggtcccg gggagccaga tgtgcgggat ggcttcagcg ccacgtttga
     1321 gaagattctg gagtcagagc tgctgcgggg cacccagtac agcagcctcg actccctaga
     1381 cgggctgagc ctcacggatg agagcgacag ctgcgtcagc ttcgaggccc ccctcacacc
     1441 cctcatccag cagcgggccc gtgacagccc tgagccaggg gctgggttgg gcattgggga
     1501 catggcgttt gagggggaca tgggggcggc tggtggtgat ggggagctgg gcagccccct
     1561 gcggcgctcc atctccagca gccgctctga gaatgtcctg agccgcctgt ctctcatggc
     1621 catgcccaat ggattccatg aagatggccc tcagggccca gggggggatg aggatgatga
     1681 tgaggaggac acggacaagt tgctgaactc agccagtgac cccagcctga aggatggcct
     1741 gtcagactca gactctgagc tcagcagctc ggaggggttg gagcctggta gtgcagaccc
     1801 tctggccaac gggtgccagg gggtcagtga agctgctcat cggctggcac gccgtctcta
     1861 ccacctcgag ggcttccagc gctgtgatgt ggcccggcag ctgggcaaga acaacgagtt
     1921 tagcaggctg gtggccgggg agtacctcag tttcttcgac ttctcgggct tgactctgga
     1981 cggagcactc agaacattct tgaaggcctt cccgctgatg ggggagacac aagagcgtga
     2041 gcgggtcctc acacacttct cccgccggta ctgccagtgc aaccctgatg acagcacttc
     2101 ggaagatggg atccacacgc tcacctgtgc cctgatgctg ctcaacacgg acctgcacgg
     2161 ccacaacatt ggcaaaaaga tgtcctgtca gcaattcatt gccaacttgg accagctgaa
     2221 tgatggccaa gactttgcca aagacctgct gaagaccctt tacaactcca tcaagaatga
     2281 aaagctggaa tgggccattg atgaggatga gctgaggaaa tccctgtctg agctggtgga
     2341 tgacaagttc gggacaggca cgaagaaggt gacgcgaatc ctggatggtg gcaacccctt
     2401 cctggatgtc ccacaggcgc tcagtgccac cacctacaag cacggcgtcc tgacccggaa
     2461 gactcacgct gacatggatg gcaagaggac gccccgtggg aggcgtggct ggaagaaatt
     2521 ctacgcagtg ctcaaaggga ccatcctgta cctgcagaag gatgagtaca ggcctgacaa
     2581 agctctatcg gagggtgacc tgaagaacgc cattcgcgtg catcacgctc tggccaccag
     2641 ggcctctgac tacagcaaga agtccaacgt gctgaagctt aagacagccg actggagggt
     2701 attcctcttc caggcaccga gcaaggaaga aatgctgtcc tggatcctca ggatcaacct
     2761 ggtggcagcc atcttctctg ccccggcctt cccagccgct gtcagctcca tgaagaagtt
     2821 ctgtcggccc ctgctgccct cctgcaccac ccgcctctgc caggaggagc aactgcggtc
     2881 tcatgagaat aagttgaggc agctgactgc ggagctggcc gaacacaggt gtcacccagt
     2941 cgagaggggc atcaagtcca aggaggccga ggagtaccgg ttgaaggagc actatctcac
     3001 cttcgagaaa agccgttatg agacctatat ccacctcctg gctatgaaaa tcaaagtggg
     3061 ctcagatgat ctggagcgga ttgaggcccg gctggccact ctggaagggg atgacccttc
     3121 tctccggaag acacattcaa gccctgccct cagccagggc catgtgactg gcagcaaaac
     3181 cacaaaggat gccactgggc ctgatactga cccagctttc ttgtacaaag tggttcgatc
     3241 tagaatggct agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     3301 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgctac
     3361 atacggaaag cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
     3421 aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat
     3481 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat
     3541 atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac
     3601 ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg
     3661 acacaaactc gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa
     3721 gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact
     3781 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     3841 ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat
     3901 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa
     3961 ataatgaatt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4021 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4081 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4141 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4201 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     4261 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4321 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4381 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4441 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4501 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4561 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4621 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4681 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4741 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4801 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4861 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4921 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4981 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5041 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5101 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5161 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5221 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5281 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5341 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5401 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5461 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5521 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5581 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5641 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5701 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5761 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5821 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5881 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5941 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6001 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6061 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6181 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6241 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6301 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6361 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6421 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7081 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8401 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatg
//
