LOCUS       sequence-217117-        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: pFlc045
FEATURES             Location/Qualifiers
     source          1..8186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     590..614
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        824..842
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     824..841
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     855..874
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1873..1891
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             1901..2611
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(1940..1961)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2201..2220)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2548..2569
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2628..2646
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     regulatory      4482..4491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4495..4629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4532..4551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4586..4605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4698..5055
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4698..4889
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4721..4741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4906..5041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5075..5092)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5075..5091)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5084..5106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5304..5759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5391..5410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5601..5622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5776..5795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5895..5917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5955..5973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6041..6145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6146..6214,6215..7006)
                     /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(6364..6383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7177..7765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7666..7685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7919..7936
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8053..8074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8089..8094,8095..8112,8113..8119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8127..8143
                     /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     8132..8154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8151..8167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8151..8167
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      601 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      661 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      721 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
      781 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
      841 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
      901 cggttctatc gattgaattc tatggttgga ccagggcttc tttttcgcca atgtaaaaag
      961 gaatatgccg tacatctttg cctttttctg caccggtttt ctaggtgcgg ttgtgggtgc
     1021 caatttcccc aacaatatcc agataggggg gttatttcca aaccaacaat cacaggaaca
     1081 tgcggctttt aggtttgctt tgtcacaact cacggagccc cccaagctgc ttccccagat
     1141 cgatattgtg aacatcagcg acagctttga gatgacttac cgtttctgtt cccagttctc
     1201 caaaggagtc tatgccatct ttggatttta tgaacgaagg actgtcaaca tgctgacctc
     1261 cttctgtggg gccctccatg tgtgcttcat tactccaagt tttcctgttg acacatccaa
     1321 tcaatttgtc cttcagctac gcccggaact acaggaagct ctcattagca ttatcgacca
     1381 ttacaagtgg caaacctttg tctacattta tgatgctgac cggggcctgt cagtcctgca
     1441 gagagtcttg gatacagccg cagagaagaa ctggcaggta acggctgtca acattctgac
     1501 aaccaccgag gaaggatacc ggatgctctt tcaggacctg gagaagaaaa aggagaggct
     1561 ggtggtggtt gactgtgaat cagaacgcct caacgccatc ctgggccaga tcgtgaagct
     1621 agaaaagaat ggcatcgggt accactacat cctcgccaat ctgggcttca tggacattga
     1681 cttaaataag ttcaaggaga gcggagccaa tgtgacaggt ttccagctgg tgaactacac
     1741 agacacgatc ccagccagaa tcatgcagca atggaggaca agtgactccc gagaccatac
     1801 cagggtggac tggaagaggc caaagtacac ttctgctctc acctatgatg gtgtcaaggt
     1861 gatggctgag gcctgtctct tatacacatc tggcgcgccg agcaagggcg aggagctgtt
     1921 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1981 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2041 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctggggcgt
     2101 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2161 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2221 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2281 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acatcagcca
     2341 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggcccact tcaagatccg
     2401 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2461 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     2521 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2581 gatcactctc ggcatggacg agctgtacaa gggcggcgcg cccgggcaga tgtgtataag
     2641 agacagggct gaggccttcc aaagcctgcg gaggcagagg attgacatat cccgccgggg
     2701 gaatgctggg gactgtctgg ctaacccagc tgtgccctgg ggtcaaggga tcgacatcca
     2761 gagagccctg cagcaggtgc gcttcgaagg tttgacagga aatgtgcagt tcaacgagaa
     2821 agggcgccgg accaattaca ccctccacgt gatcgaaatg aaacatgatg gaatccgaaa
     2881 gattggttac tggaatgaag acgataaatt tgtccccgca gccaccgacg ctcaggctgg
     2941 aggggacaac tcaagcgtcc agaataggac ctacatcgtc actactatcc tcgaagatcc
     3001 ttacgtgatg cttaaaaaga atgccaacca gtttgagggc aatgaccgct atgagggcta
     3061 ctgtgtggag ctggctgcag agatcgccaa gcacgtgggc tactcctacc gacttgagat
     3121 tgtcagcgac ggcaaatatg gagcccggga tcccgacaca aaggcttgga atggcatggt
     3181 gggagaactg gtctatggaa gagcagacgt ggctgtggct cccttgacca taaccttggt
     3241 ccgggaggaa gtcatcgact tctccaagcc attcatgagt ttgggaatct ccattatgat
     3301 taagaagcca cagaagtcca agccaggtgt cttctccttt cttgaccctt tggcctatga
     3361 gatctggatg tgtatagtgt ttgcctacat tggagtgagc gtcgtcctct tcctggtcag
     3421 ccgtttcagc ccctacgaat ggcacagcga agagtttgaa gagggacgag accagacaac
     3481 cagtgaccag tcaaatgagt ttggcatatt caacagcctg tggttctccc tgggggcctt
     3541 catgcagcaa ggatgtgaca tttcccccag gtccctgtcc ggacgcatcg tcggcggcgt
     3601 ctggtggttc ttcactttga tcatcatctc ctcgtacaca gccaacctgg ctgccttcct
     3661 gactgtggag aggatggtgt ctcccattga gagtgcagag gacctggcaa agcagacgga
     3721 aattgcttat gggacattgg aagcaggctc cactaaggag ttcttcagga gatctaaaat
     3781 cgctgtgttt gagaagatgt ggacatacat gaagtctgca gaaccatccg tgtttgttcg
     3841 gaccacagag gaaggcatga tcagagtgag aaaatctaaa ggcaaatacg cctacctcct
     3901 ggagtccacc atgaatgagt atattgagca acgaaagccc tgtgacacca tgaaagtggg
     3961 aggtaacttg gattccaaag gctatggcat tgcgacaccc aaggggtccg ccctgagagg
     4021 tcccgtaaac ctagcggttt tgaaactcag tgagcaaggc gtcttagaca agctgaaaag
     4081 caaatggtgg tacgataaag gggaatgtgg aagcaaggac tccggaagta aggacaagac
     4141 cagcgctttg agcctcagca atgtggcagg cgtgttctac atcctgattg gagggctggg
     4201 actggccatg ctggttgcct taatcgagtt ctgctacaaa tcccgtagcg agtcgaagcg
     4261 gatgaagggt ttctgtttga tcccacagca atccatcaat gaagccatac ggacatcgac
     4321 cctcccccgg aacagtgggg caggagccag cggaggaggc ggcagtggag agaatggccg
     4381 ggtggtcagc caggacttcc ccaagtccat gcaatccatt ccctgcatga gtcacagttc
     4441 agggatgccc ttgggagcca caggattgta actgaagctt ggccgccatg gcccaacttg
     4501 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4561 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4621 gtctggatcg ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga
     4681 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4741 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4801 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4861 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4921 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4981 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5041 ctaggctttt gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg
     5101 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     5161 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     5221 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5281 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5461 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5641 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5701 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     5761 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5821 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5881 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5941 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     6001 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6061 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6121 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6181 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6241 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6301 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6361 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6421 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6481 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6541 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6601 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6661 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6721 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6781 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6841 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6901 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6961 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7021 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7081 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7141 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7201 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7261 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7321 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7381 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7441 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7501 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7561 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7621 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7681 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7741 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7801 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7861 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7921 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7981 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     8041 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     8101 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     8161 ctatgacatg attacgaatt aattcg
//
