LOCUS       sequence-217095-        8186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent, functional, glutamate receptor subunit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8186)
  AUTHORS   Sheridan DL, Robert A, Cho CH, Howe JR, Hughes TE
  TITLE     Regions of alpha-amino-5-methyl-3-hydroxy-4-isoxazole propionic acid
            receptor subunits that are permissive for the insertion of green 
            fluorescent protein.
  JOURNAL   Neuroscience. 2006 Aug 25;141(2):837-49. Epub 2006 Jun 12.
  PUBMED    16765522
REFERENCE   2  (bases 1 to 8186)
  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: 
            "Neuroscience."; date: "2006-08-25"; volume: "141(2)"; pages: 
            "837-49. Epub 2006 Jun 12"
COMMENT     Sequence Label: pFlg262
FEATURES             Location/Qualifiers
     source          1..8186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1806..1824
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             1833..2543
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             1833..2543
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(1872..1893)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2133..2152)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2480..2501
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2561..2579
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     regulatory      4486..4495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4499..4633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4536..4555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4590..4609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4702..5059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4702..4893
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4725..4745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4910..5045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5079..5096)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5079..5095)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5088..5110)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5308..5763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5395..5414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5605..5626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5780..5799)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5899..5921
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5959..5977)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6045..6149
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6150..6218,6219..7010)
                     /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(6368..6387)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7181..7769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7670..7689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7923..7940
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8057..8078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8093..8098,8099..8116,8117..8123)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8131..8147
                     /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     8136..8158
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8155..8171
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8155..8171
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attctatggt tggaccaggg cttctttttc gccaatgtaa
      961 aaaggaatat gccgtacatc tttgcctttt tctgcaccgg ttttctaggt gcggttgtgg
     1021 gtgccaattt ccccaacaat atccagatag gggggttatt tccaaaccaa caatcacagg
     1081 aacatgcggc ttttaggttt gctttgtcac aactcacgga gccccccaag ctgcttcccc
     1141 agatcgatat tgtgaacatc agcgacagct ttgagatgac ttaccgtttc tgttcccagt
     1201 tctccaaagg agtctatgcc atctttggat tttatgaacg aaggactgtc aacatgctga
     1261 cctccttctg tggggccctc catgtgtgct tcattactcc aagttttcct gttgacacat
     1321 ccaatcaatt tgtccttcag ctacgcccgg aactacagga agctctcatt agcattatcg
     1381 accattacaa gtggcaaacc tttgtctaca tttatgatgc tgaccggggc ctgtcagtcc
     1441 tgcagagagt cttggataca gccgcagaga agaactggca ggtaacggct gtcaacattc
     1501 tgacaaccac cgaggaagga taccggatgc tctttcagga cctggagaag aaaaaggaga
     1561 ggctggtggt ggttgactgt gaatcagaac gcctcaacgc catcctgggc cagatcgtga
     1621 agctagaaaa gaatggcatc gggtaccact acatcctcgc caatctgggc ttcatggaca
     1681 ttgacttaaa taagttcaag gagagcggag ccaatgtgac aggtttccag ctggtgaact
     1741 acacagacac gatcccagcc agaatcatgc agcaatggag gacaagtgac tcccgagacc
     1801 ataccctgtc tcttatacac atctggcgcg ccagcaaggg cgaggagctg ttcaccgggg
     1861 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1921 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1981 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2041 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2101 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2161 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2221 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2281 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2341 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2401 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2461 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2521 tcggcatgga cgagctgtac aagggtgggc gcgcccgggc agatgtgtat aagagacagg
     2581 accataccag ggtggactgg aagaggccaa agtacacttc tgctctcacc tatgatggtg
     2641 tcaaggtgat ggctgaggcc ttccaaagcc tgcggaggca gaggattgac atatcccgcc
     2701 gggggaatgc tggggactgt ctggctaacc cagctgtgcc ctggggtcaa gggatcgaca
     2761 tccagagagc cctgcagcag gtgcgcttcg aaggtttgac aggaaatgtg cagttcaacg
     2821 agaaagggcg ccggaccaat tacaccctcc acgtgatcga aatgaaacat gatggaatcc
     2881 gaaagattgg ttactggaat gaagacgata aatttgtccc cgcagccacc gacgctcagg
     2941 ctggagggga caactcaagc gtccagaata ggacctacat cgtcactact atcctcgaag
     3001 atccttacgt gatgcttaaa aagaatgcca accagtttga gggcaatgac cgctatgagg
     3061 gctactgtgt ggagctggct gcagagatcg ccaagcacgt gggctactcc taccgacttg
     3121 agattgtcag cgacggcaaa tatggagccc gggatcccga cacaaaggct tggaatggca
     3181 tggtgggaga actggtctat ggaagagcag acgtggctgt ggctcccttg accataacct
     3241 tggtccggga ggaagtcatc gacttctcca agccattcat gagtttggga atctccatta
     3301 tgattaagaa gccacagaag tccaagccag gtgtcttctc ctttcttgac cctttggcct
     3361 atgagatctg gatgtgtata gtgtttgcct acattggagt gagcgtcgtc ctcttcctgg
     3421 tcagccgttt cagcccctac gaatggcaca gcgaagagtt tgaagaggga cgagaccaga
     3481 caaccagtga ccagtcaaat gagtttggca tattcaacag cctgtggttc tccctggggg
     3541 ccttcatgca gcaaggatgt gacatttccc ccaggtccct gtccggacgc atcgtcggcg
     3601 gcgtctggtg gttcttcact ttgatcatca tctcctcgta cacagccaac ctggctgcct
     3661 tcctgactgt ggagaggatg gtgtctccca ttgagagtgc agaggacctg gcaaagcaga
     3721 cggaaattgc ttatgggaca ttggaagcag gctccactaa ggagttcttc aggagatcta
     3781 aaatcgctgt gtttgagaag atgtggacat acatgaagtc tgcagaacca tccgtgtttg
     3841 ttcggaccac agaggaaggc atgatcagag tgagaaaatc taaaggcaaa tacgcctacc
     3901 tcctggagtc caccatgaat gagtatattg agcaacgaaa gccctgtgac accatgaaag
     3961 tgggaggtaa cttggattcc aaaggctatg gcattgcgac acccaagggg tccgccctga
     4021 gaggtcccgt aaacctagcg gttttgaaac tcagtgagca aggcgtctta gacaagctga
     4081 aaagcaaatg gtggtacgat aaaggggaat gtggaagcaa ggactccgga agtaaggaca
     4141 agaccagcgc tttgagcctc agcaatgtgg caggcgtgtt ctacatcctg attggagggc
     4201 tgggactggc catgctggtt gccttaatcg agttctgcta caaatcccgt agcgagtcga
     4261 agcggatgaa gggtttctgt ttgatcccac agcaatccat caatgaagcc atacggacat
     4321 cgaccctccc ccggaacagt ggggcaggag ccagcggagg aggcggcagt ggagagaatg
     4381 gccgggtggt cagccaggac ttccccaagt ccatgcaatc cattccctgc atgagtcaca
     4441 gttcagggat gcccttggga gccacaggat tgtaactgaa gcttggccgc catggcccaa
     4501 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4561 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4621 tcatgtctgg atcgggaatt aattcggcgc agcaccatgg cctgaaataa cctctgaaag
     4681 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4741 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4801 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4861 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4921 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4981 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     5041 aggcctaggc ttttgcaaaa agctgttaac agcttggcac tggccgtcgt tttacaacgt
     5101 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5161 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5221 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5281 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     5341 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5401 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5461 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5521 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5581 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5641 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5701 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     5761 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5821 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5881 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5941 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6001 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6061 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6121 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6181 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6241 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6301 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6361 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6421 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6481 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6541 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6601 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6661 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6721 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6781 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6841 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6901 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6961 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7021 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7321 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7801 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7861 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7921 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7981 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     8041 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     8101 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     8161 acagctatga catgattacg aattaa
//
