LOCUS       sequence-217122-        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: pFlc245
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    3629..3647
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             3657..4367
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(3696..3717)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3957..3976)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4304..4325
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4384..4402
                     /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 ataccagggt ggactggaag aggccaaagt acacttctgc tctcacctat gatggtgtca
     1861 aggtgatggc tgaggccttc caaagcctgc ggaggcagag gattgacata tcccgccggg
     1921 ggaatgctgg ggactgtctg gctaacccag ctgtgccctg gggtcaaggg atcgacatcc
     1981 agagagccct gcagcaggtg cgcttcgaag gtttgacagg aaatgtgcag ttcaacgaga
     2041 aagggcgccg gaccaattac accctccacg tgatcgaaat gaaacatgat ggaatccgaa
     2101 agattggtta ctggaatgaa gacgataaat ttgtccccgc agccaccgac gctcaggctg
     2161 gaggggacaa ctcaagcgtc cagaatagga cctacatcgt cactactatc ctcgaagatc
     2221 cttacgtgat gcttaaaaag aatgccaacc agtttgaggg caatgaccgc tatgagggct
     2281 actgtgtgga gctggctgca gagatcgcca agcacgtggg ctactcctac cgacttgaga
     2341 ttgtcagcga cggcaaatat ggagcccggg atcccgacac aaaggcttgg aatggcatgg
     2401 tgggagaact ggtctatgga agagcagacg tggctgtggc tcccttgacc ataaccttgg
     2461 tccgggagga agtcatcgac ttctccaagc cattcatgag tttgggaatc tccattatga
     2521 ttaagaagcc acagaagtcc aagccaggtg tcttctcctt tcttgaccct ttggcctatg
     2581 agatctggat gtgtatagtg tttgcctaca ttggagtgag cgtcgtcctc ttcctggtca
     2641 gccgtttcag cccctacgaa tggcacagcg aagagtttga agagggacga gaccagacaa
     2701 ccagtgacca gtcaaatgag tttggcatat tcaacagcct gtggttctcc ctgggggcct
     2761 tcatgcagca aggatgtgac atttccccca ggtccctgtc cggacgcatc gtcggcggcg
     2821 tctggtggtt cttcactttg atcatcatct cctcgtacac agccaacctg gctgccttcc
     2881 tgactgtgga gaggatggtg tctcccattg agagtgcaga ggacctggca aagcagacgg
     2941 aaattgctta tgggacattg gaagcaggct ccactaagga gttcttcagg agatctaaaa
     3001 tcgctgtgtt tgagaagatg tggacataca tgaagtctgc agaaccatcc gtgtttgttc
     3061 ggaccacaga ggaaggcatg atcagagtga gaaaatctaa aggcaaatac gcctacctcc
     3121 tggagtccac catgaatgag tatattgagc aacgaaagcc ctgtgacacc atgaaagtgg
     3181 gaggtaactt ggattccaaa ggctatggca ttgcgacacc caaggggtcc gccctgagag
     3241 gtcccgtaaa cctagcggtt ttgaaactca gtgagcaagg cgtcttagac aagctgaaaa
     3301 gcaaatggtg gtacgataaa ggggaatgtg gaagcaagga ctccggaagt aaggacaaga
     3361 ccagcgcttt gagcctcagc aatgtggcag gcgtgttcta catcctgatt ggagggctgg
     3421 gactggccat gctggttgcc ttaatcgagt tctgctacaa atcccgtagc gagtcgaagc
     3481 ggatgaaggg tttctgtttg atcccacagc aatccatcaa tgaagccata cggacatcga
     3541 ccctcccccg gaacagtggg gcaggagcca gcggaggagg cggcagtgga gagaatggcc
     3601 gggtggtcag ccaggacttc cccaagtcct gtctcttata cacatctggc gcgccgagca
     3661 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3721 acggccacaa gttcagtgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     3781 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3841 ccctgacctg gggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     3901 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3961 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4021 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4081 acaactacat cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     4141 cccacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     4201 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     4261 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4321 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggcgcgcccg
     4381 ggcagatgtg tataagagac agccccaagt 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
//
