LOCUS       sequence-233694-        9695 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 9695)
  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 9695)
  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-NR2b wt
FEATURES             Location/Qualifiers
     source          1..9695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1197..1199,1200..1910)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1197..1199,1200..1910)
                     /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(1239..1260)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1500..1519)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1847..1868
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6809..6930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6820..6839)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6874..6893
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      7111..7566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7198..7217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7408..7429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7633..7652)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7752..7774
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7812..7830)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7898..8002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8003..8071,8072..8863)
                     /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(8221..8240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9034..9622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9523..9542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagaggt ttggcgtcta caaaccaaga
     1081 gagccgacta gctgaagatg aagcccagcg cagagtgctg ttcccccaag ttctggttgg
     1141 tgttggccgt cttggccgta tcaggcagca aagctcgttc ccaaaagagc gctagggtga
     1201 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1261 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1321 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1381 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1441 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1501 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1561 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1621 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1681 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1741 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1801 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1861 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag gctagccccc
     1921 ccagcatcgg catcgctgtc atcctcgtgg gcacttcaga cgaagtggcc ataaaagacg
     1981 cccacgagaa agatgacttc catcatctct cagtagttcc ccgggtggag ctggtagcca
     2041 tgaacgaaac tgacccaaag agcatcatca cccgtatctg cgatcttatg tctgaccgga
     2101 agatccaggg ggtggtgttc gcggatgaca ccgaccaaga agccatcgct cagatcctcg
     2161 acttcatttc tgctcagact ctcaccccca tcctgggcat ccatgggggc tcatctatga
     2221 taatggcgga taaggatgag tcctccatgt tcttccagtt tggcccgtct atcgaacagc
     2281 aagcttccgt catgctcaac atcatggaag aatatgactg gtacatcttt tccatcgtca
     2341 ccacctactt ccctggctac caggactttg tgaacaagat ccgcagtacc atcgagaaca
     2401 gcttcgtggg ctgggagctc gaggaagtcc tcctgctaga catgtctctg gacgatggcg
     2461 actctaagat tcagaatcag ctgaagaagc tccaaagccc catcattctc ctttattgca
     2521 cgaaggagga agccacctac atttttgaag tagctaactc agttgggctg actggctacg
     2581 gctacacgtg gattgtgccg agtctggtgg ccggggatac ggacacggtg ccttcagagt
     2641 tccccacggg gcttatctct gtgtcttatg atgaatggga ctatggcctt cctgccagag
     2701 tgagagatgg aattgccatc atcaccactg ctgcctcgga catgctgtcc gaacacagtt
     2761 tcatccctga gcccaagagc agttgctaca acacccacga gaagaggatc taccagtcta
     2821 acatgttgaa taggtatctg atcaatgtca cttttgaagg gagaaacctg tccttcagcg
     2881 aagatggcta ccagatgcat ccgaagctgg tgataatcct tctgaacaag gagaggaagt
     2941 gggagagggt ggggaaatgg aaggacaagt ccctgcagat gaagtattat gtgtggcctc
     3001 ggatgtgtcc tgagactgag gagcaagagg atgaccatct gagcattgtc accttggagg
     3061 aggcgccatt tgtcattgtg gaaagcgtgg accctctcag tggaacctgc atgaggaata
     3121 cagtcccgtg ccagaagcgc atcatctctg agaataaaac agatgaggaa ccaggctaca
     3181 tcaaaaaatg ctgcaagggg ttctgtattg acatccttaa gaaaatttct aagtctgtga
     3241 agttcaccta tgacctttac ctggtgacca atggcaagca cgggaagaag attaatggga
     3301 cctggaatgg catgatcggt gaggtggtcc tgaagagggc ctacatggca gtgggatcac
     3361 taactatcaa tgaagaacgg tcagaggtgg ttgacttctc tgtacccttc atagaaactg
     3421 gcatcagtgt catggtatct cgcagcaatg ggactgtgtc accttctgcc ttcttagagc
     3481 cattcagcgc tgacgtgtgg gtgatgatgt ttgtgatgct gctcattgtt tctgcggtgg
     3541 ctgtctttgt ctttgaatac ttcagccctg tgggttacaa caggtgccta gccgatggca
     3601 gagagccagg aggcccatct ttcaccatcg gcaaagcaat ttggttactc tggggtctgg
     3661 tgtttaacaa ctccgtacct gtgcagaacc caaaggggac cacctccaag atcatggtgt
     3721 cagtgtgggc cttctttgct gtcattttcc tggccagcta cactgccaac ttagcagcct
     3781 tcatgatcca agaggagtat gtggaccagg tttctggcct gagtgacaag aagttccaga
     3841 gacctaatga cttctcaccc cctttccgct ttgggactgt gcccaatggc agcacagaga
     3901 ggaatatccg taataactat gcagaaatgc atgcctacat gggaaagttc aaccaaaggg
     3961 gtgtagatga tgcattgctc tccctgaaaa cagggaagct tgatgcattc atctatgatg
     4021 cagctgtgct caactacatg gctggaaggg acgaaggctg caaactggtg accattggca
     4081 gtggcaaggt ctttgcttct accggctatg gcattgctat ccaaaaggac tccgggtgga
     4141 agcgccaggt ggacctggct atcctgcagc tgtttggaga tggggagatg gaagaactgg
     4201 aagctctctg gctcactggc atttgccaca atgagaagaa tgaggtgatg agcagccagc
     4261 tggacatcga caatatggca ggtgtcttct atatgttggg ggcagccatg gccctcagcc
     4321 tcatcacctt catctgtgag catctgttct attggcagtt ccggcattgc ttcatgggtg
     4381 tctgttctgg caagcctggc atggtcttct ccatcagcag aggtatctac agctgtatcc
     4441 atggggtagc catagaggag cgccaatccg tgatgaactc ccccactgcc accatgaaca
     4501 acacccactc caacatccta cgcttgctcc gcacggccaa gaacatggcc aacctgtctg
     4561 gagtaaacgg ctcccctcag agtgccctgg acttcatccg ccgagagtcc tccgtctacg
     4621 acatctctga gcatcgtcgc agcttcacgc attcagactg caagtcttac aataacccac
     4681 cctgtgagga aaacctgttc agtgactaca ttagcgaggt agagagaaca tttggtaacc
     4741 tgcagctgaa ggacagcaat gtgtaccaag accactatca ccatcaccac cggccacaca
     4801 gcatcggcag caccagctcc attgatgggc tctatgactg tgacaaccca cccttcacca
     4861 cccagcccag gtcaatcagc aagaaacccc tggacatcgg cctgccctcc tccaaacata
     4921 gccagctcag cgacctgtat ggcaagttct ctttcaagag tgaccgctac agtggccacg
     4981 acgacttgat tcgatcggat gtctccgaca tctccacgca cactgtcacc tatgggaaca
     5041 tcgagggcaa cgcagccaag aggaggaaac agcagtataa ggacagtcta aagaagcggc
     5101 cagcctcggc caaatcgagg agggagtttg atgaaatcga gctggcctac cgtcgccgac
     5161 caccccgctc cccggaccac aagcgctact tcagggacaa agaagggctc cgagacttct
     5221 acctggacca gttccgaaca aaggagaact cgcctcactg ggagcacgtg gacttgactg
     5281 acatttacaa agaacgcagt gacgacttca agcgagattc ggtcagtgga ggtgggccct
     5341 gtaccaacag gtctcacctc aaacacggaa cgggcgagaa gcacggagtg gtaggcgggg
     5401 tgcctgctcc ttgggagaag aacctgacca atgtggattg ggaggaccgg tctgggggca
     5461 acttctgccg cagctgtcct tccaagctgc acaattactc ctcgacggtg gcagggcaga
     5521 actcgggccg gcaggcctgc atcagatgtg aggcctgtaa gaaggctggt aacctgtacg
     5581 acatcagcga ggacaactcc ctgcaggaac tggaccagcc ggctgccccc gtggctgtga
     5641 catccaacgc ctccagcacc aagtaccctc aaagcccgac taattccaag gctcagaaga
     5701 agaatcggaa caaactgcgc cggcagcatt cctacgacac cttcgtggac ctgcagaagg
     5761 aggaggccgc cttggcccca cgcagcgtga gcctgaaaga caagggccga ttcatggatg
     5821 ggagccccta cgcccatatg tttgagatgc cagctggtga gagctccttt gccaacaagt
     5881 cctcagtgcc cactgccgga caccaccaca acaaccccgg cagcggctac atgctcagca
     5941 agtcgctcta ccctgaccgg gtcacgcaaa accctttcat ccccactttt ggggatgacc
     6001 agtgcttgct tcacggcagc aaatcctact tcttcaggca gcccacggtg gcaggggcgt
     6061 caaaaacaag gccggacttc cgggcccttg tcaccaataa gccagtggtg tcagcccttc
     6121 atggggctgt gccaggtcgt ttccagaagg acatttgtat agggaaccag tccaacccct
     6181 gtgtgcctaa caacaaaaac cccagggctt tcaatggctc cagcaatgga catgtttatg
     6241 agaaactttc tagtattgag tctgatgtct gagtgaggga agagagagag gttaaggtgg
     6301 gtacgggagg gatagggctg tgggccgcgt ggtgcgcatg tcatggaaag agtcgggggt
     6361 gaacttggct cccattcgct ttttcttctt cttttaattt ctctatggga tcctggagtt
     6421 ctggttcctt actgaaggca accctcgtgg ccagcaccat ttctcctccc tcgcgcagtt
     6481 ctctccttcc cgcatctgtc caccattcct gtttccatga gagaatagaa cggggcctca
     6541 gtgtgggagg atggagagga gaccaaagcg ctgcctgtgt gcttctttct agcgcagaag
     6601 ggctcttagc agttcacttt gagcgaggct ttcctgatgc tgctctcttt gttcaggtga
     6661 ggaagcgaag gtgttctgag gaaggccatt gaacctcctg ttcataagag agaagaggct
     6721 ttcactgaat tcttgccgcg tggttgacct cgaggggggg cccggtacct ctagagtcga
     6781 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     6841 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     6901 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     6961 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     7021 aaaatcgata aggatccggg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     7081 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     7141 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     7201 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     7261 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     7321 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     7381 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     7441 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     7501 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     7561 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     7621 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     7681 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     7741 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     7801 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7861 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7981 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     8041 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     8101 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     8161 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     8221 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     8281 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     8341 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     8401 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     8461 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     8521 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     8581 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     8641 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8701 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     8761 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8821 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8881 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8941 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     9001 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     9061 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     9121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     9181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     9241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     9301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     9661 ttgctggcct tttgctcaca tggctcgaca gatct
//
