LOCUS       sequence-364538-        8994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8994)
  AUTHORS   Barria A, Malinow R
  TITLE     Subunit-specific NMDA receptor trafficking to synapses.
  JOURNAL   Neuron. 2002 Jul 18;35(2):345-53.
  PUBMED    12160751
REFERENCE   2  (bases 1 to 8994)
  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."; 
            date: "2002-07-18"; volume: "35(2)"; pages: "345-53"
COMMENT     Sequence Label: pCI-EGFP-NR1 wt
FEATURES             Location/Qualifiers
     source          1..8994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..198
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     438..459
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(914..932)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(975..994)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          977..1109
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     complement(1224..1248)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1245..1448)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1274..1294)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1449..1828
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2039..2627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2119..2138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2798..3589,3590..3658))
                     /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     3421..3440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3659..3763)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3831..3849
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3887..3909)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4009..4028
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4095..4550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4232..4253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4444..4463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4731..4852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4768..4787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4822..4841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8782..8994,1..498,499..501))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(8782..8994,1..498,499..501))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8824..8845)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
       61 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
      121 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
      181 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
      241 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
      301 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
      361 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
      421 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
      481 cagctcctcg cccttgctca ccctagtggc ggcgcgggcg aaggagcagg aaaaaagcag
      541 ggcgaatgtc agcaggtgca tggtgctcat gagctccggg cacagcgggc ctggctctgg
      601 gttgcgcgag tgcgacgacc tcgcgatgta cgacccgcgg gccgcccgcg cgcggtcaag
      661 cgcccccggg ttctctagga gccccgggcg agcgcggcgg atgctgtccg aggtgctgaa
      721 gcgtgtttgg cgctgctcga acacccgcgc ggaaaaggcg gaaaggacag cagcgcggac
      781 tgcgcgactg gtctcttaga aacctctaca gaaactgcct taacaaacca atattcgaca
      841 gcaacggaat tcgatatcaa gcttatcgat accgtcgact ctagaggtac cacgcgtgaa
      901 ttctcgaggc tagcctatag tgagtcgtat taagtactct agccttaaga gctgtaattg
      961 aactgggagt ggacacctgt ggagagaaag gcaaagtgga tgtcagtaag accaataggt
     1021 gcctatcaga aacgcaagag tcttctctgt ctcgacaagc ccagtttcta ttggtctcct
     1081 taaacctgtc ttgtaacctt gatacttacc tgcccagtgc ctcacgacca acttctgcag
     1141 cttaagttcg agactgttgt gtcagaagca ctgactgcgt tagcaattta actgtgataa
     1201 actaccgcaa taaagcttct agtgatctga cggttcacta aacgagctct gcttatatag
     1261 acctcccacc gtacacgcct accgcccatt tgcgtcaacg gggcggggtt attacgacat
     1321 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     1381 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattggtgta ctgccaaaac
     1441 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     1501 cgtaaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     1561 gggggcggac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     1621 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     1681 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     1741 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     1801 tattaataac tagtcaataa tcaatgccaa catggcggtc atattggaca tgagccaata
     1861 taaatgtaca tattatgata tagatacaac gtatgcaatg gccaatagcc aatattgatt
     1921 tatgctatat aaccaatgaa taatatggct aatggccaat attgaagatc tgtcgagcca
     1981 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2041 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2101 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2161 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2221 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2281 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2341 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2401 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2461 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2521 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2581 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2641 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2701 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2761 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2821 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2881 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2941 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3001 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3061 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3121 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3181 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3241 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3301 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3361 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3421 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3481 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3541 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3601 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3661 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3721 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3781 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3841 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3901 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3961 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4021 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4081 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4141 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4201 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4261 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4321 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4381 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4441 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4501 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     4561 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4621 cccggatcct tatcgatttt accacatttg tagaggtttt acttgcttta aaaaacctcc
     4681 cacatctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4741 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4801 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4861 gctcgaagcg gccgcttttt tttttttttt ctgacattcc tttaggcatc accactgaat
     4921 accgggatta ggggtggctg tgcgcgcccg gtgtccctgg ggcagacact gggacactgg
     4981 gagagctggt catgtacaag gtagtggggt gagtgacacc ggactgggag gcagatggcg
     5041 ctccctggtg cccaaggtgc gggactcatc acgagccagc gtctgaggaa gcctattgtg
     5101 gctgcggaca gaagagaagg ctcagccctc ttggagtctg gagccctaga tgtctgggag
     5161 ccagaaagca cccctgaagc actgccacta gggcagtgaa ggctagccct gggaaaggtc
     5221 agagacaacc cagtgaatgg ggtggggaaa tgtcgtggga gggtggtgca gcaggagaga
     5281 ggtggaaatc aagttgcaga gagcaggcgc tggaggaagg gtcgggggtg acctagcaag
     5341 ctggggccat aggcccccac tgtagcttgg ggtagtggct ggaggaagga tggggctgcc
     5401 atttctgctc cagactcctc cgaccggggc cacttaggcc cctcacccca ctgctcagag
     5461 acctcagggc cctgcctctg ccggagcgcc ctgtggccct gcccagccgg ctcagctctg
     5521 gcccatcctc agccgctctg gtccgcacct tggtcctgca ggggctgccc gccaggcagc
     5581 agagcaggcg gacgggaggc ccagggtggg cagtctctgg ggcgggtggg cttccctcag
     5641 gccttcccga gcagcaggac tcatcagtgt ccttggccgg gctaactggg caggtgggcg
     5701 cctccggggg gcctcacacc acggtgctga ccgagggatc tgagaggttg agcgggcccg
     5761 tgatatcagt gggatggtac tgctgtaaaa tagaaataca cagacaaggc ggcccgttag
     5821 ctccgacgtc gggggcacgc gcggggccgg gacagggtgg accggccagc cggccgaccg
     5881 gtgggcaggc gggcacggcc tgggggaggc tgggagtgct cctcccccga ccccgtcgtg
     5941 ctccggggct ctgcgtgggc caggccgctg tcccgtgcgt ctgtctgcgg gggaggggca
     6001 gaggagggcg ggcggggcgt ctcagctctc cctatgacgg gaacacagct gcagctggcc
     6061 ctcctccctc tcaatagcgc gtcgcggcag cactgtgtct ttttggtttt gcaaagcgcc
     6121 gcgtccaccc ccggtgctcg tgtctttgga ggacctacgt ctcttgaagc tggaggccag
     6181 ggtggaggtg atagccctaa atgtggcttt ctttttaggg tcgggctctg ctctaccact
     6241 ctttctatcc tgcaggttct tcctccacac gttcacggct gcaaaagcca gctgcatctg
     6301 cttcctacgg gcatccttgt gtcgcttgta ggcgatctca atgaaaatga ggaaaatccc
     6361 agctacgatg cctccagcca ccagcatgaa gacccctgcc atgttctcaa aagtgagggt
     6421 tgcaggagca ttgctgcggg agtcgcattc ctgataccga acccatgtct tatccagatc
     6481 ttccatgaag ccattctcat gggacttgag tatggacagg gaaacgttct gcttccaggg
     6541 gctgtccttg cgcatgccga tgccaaagcc tgagcggaag aacagctcac ccgtggtcac
     6601 cagatcgcac ttctgtgaag cctcaaactc cagcacggcc gagtcccaga taaaggcgtg
     6661 cagcttgttg tcccgcacag cctggatggc ctcagctgcg ctctcgtaat tgtgtttttc
     6721 catgtgccgg tacatggtac tcaactccac ctgcctccgg aagtagatgt ccacggagct
     6781 ctgctttaca gttgcgtaga tgaacttgtc tgaggggttt ctgagcctgg ggtcattgat
     6841 gcccgtgatg cgctcctcag gccgatccag caccaggaaa gctgccaagt tggcagtgta
     6901 ggaagccact atgatcatgg cgaaaccagc ccacaccatg cctaggatac gtgcagagaa
     6961 actccggggg gcaccttccc caatgccgga gttgagcagg acgccccagg aaaaccacat
     7021 ggcagaggac agggtcagtg catcttcctc ctcctcctca ctgttcacct tgaatcggcc
     7081 aaagggactg aagcggtcca gcaggtacag catcacagcc accacatgaa ctgacagtcc
     7141 tactagcaac cacagtgtgc tctgaaaagg ctgcataaat gagtccagtg tgctcctggg
     7201 aatctccttc ttgaccaaaa tggtcaggcc ctggtacttg aagggcttgg agaactctat
     7261 gtactgcgca cgctcattgt tgatggtcag tggtgccaca atcatgtccg cttggccact
     7321 gagtagctcg cccatcattc cgttccactc ctttttgttg ctgttgttta cccgctcctg
     7381 tgtgccaaac ttgccatctg ccaccaggtg cacctcatag gtaaaattca tggtccgcgc
     7441 cagcttgatg agcaggtcta tgcagaagcc atagcagcac tggggcactg tgtggcgtgg
     7501 gctgcctggg gacgtgtcat taggccccgt acagatcacc ttcttcactg ggtcaccatt
     7561 gactgtgaac tcctctttgc atgtcccatc actcattgtg ggcttgacgt acacgaaggg
     7621 ctcttggtgg attgtcacta tctttagtct ggtggacatc tggtatcctc gaggtttctc
     7681 tgtctctcct cctggccaga tgatcttcct gtcatttggg atgacatggg taccattgta
     7741 gatgcccact tgcaccagct tgcggttctg caggttcatg atactatagt tggcaaactt
     7801 ccggtcccca tcctcattga attccacacg gccagtcact ccgtccgcat acttagaaga
     7861 catcagcacc ctcttgaaca atggtcctgt cttccagatg ttggtgttgc ccacgcaacc
     7921 ccgcggtggg tcagtgatat tctccttctc taggagttcg tgaactgcct gtgccaccac
     7981 gcccacggcg tcactgatgt gggctgactc attcttgcca ttgatgagct gaagtccgat
     8041 gatgccatca ggagcgtagc gcagggcgtt cccagagatc tcgcgttccc cgaccagcca
     8101 cacgtaccca gagcccgtca tgttcagcat tgcggctgcg cggtacactg tggcagcgtc
     8161 gtcctcgctt gcagaaagga tgatgacccg ggcctccagt tcccgggcct ccatcagcag
     8221 agccgtcaca ttcttggttc ctgggtcaaa ctgcagcacc ttctctgcct tggactcccg
     8281 ttcctccagc aacgtctcca agcgcttctg cgctgcccgt ccctcgtggt cgtcgctgac
     8341 cagcaggatg atgtggttcc agttgtagac tcgcatcatc tcaaaccaga cgctggactg
     8401 gtgggagtag ggcggcaccg tgcgaaggaa actcaggtgg atactcttgt cagagtagat
     8461 ggacattcgg gtagtcagtc ccaggacagg gattctgtag aagccagctg tgtaggagac
     8521 aggggtggga gtgaagtggt cgttgggagt aggcgggtgg ctaactagga tagcgtagac
     8581 ctggctagag atgaggtcct cacacactga cagggccatc tgtatggcgt tgggcttgtg
     8641 ggtgacagaa gtggcgttga gctgtatctt ccaagagccg tgtcgcttat tggcctggtt
     8701 tactgcctcg cggaacatct gttcatgctt gcgcgtgctc agcaccgcgc cgatgttgac
     8761 gatcttgggg tcgcaactag ccttgtacag ctcgtccatg ccgagagtga tcccggcggc
     8821 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     8881 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     8941 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcg
//
