LOCUS       sequence-107617-        7889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human duplicated Alpha7 (CHRFAM7A) with 
            mEGFP fused into M3-M4 loop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  AUTHORS   Wang Y, Xiao C, Indersmitten T, Freedman R, Leonard S, Lester HA
  TITLE     The duplicated alpha7 subunits assemble and form functional 
            nicotinic receptors with the full-length alpha7.
  JOURNAL   J Biol Chem. 2014 Sep 19;289(38):26451-63. doi: 
            10.1074/jbc.M114.582858. Epub 2014 Jul 23.
  PUBMED    25056953
REFERENCE   2  (bases 1 to 7889)
  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"; doi: 
            "10.1074/jbc.M114.582858"; journalName: "J Biol Chem"; date: 
            "2014-09-19- 19"; volume: "289"; issue: "38"; pages: "26451-63"
COMMENT     Sequence Label: pcDNA3.1-CHRFAM7A-mGFP
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2364..2366,2367..2369,2370..3080)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2409..2430)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2670..2689)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3017..3038
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3483..3500)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3489..3713
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3759..4187
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3846..3865)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4056..4077
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4196..4216)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4201..4530
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4381..4516
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4443..4462
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4597..5391
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4651..4670)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5261..5280
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5565..5686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5602..5621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5656..5675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5735..5751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5735..5751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5748..5770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5759..5775
                     /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)."
     promoter        complement(join(5783..5789,5790..5807,5808..5813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5828..5849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5966..5983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6137..6722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6217..6236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6893..7684,7685..7753))
                     /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     7516..7535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7754..7858)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattgccc
      961 ttctcggtgc cccttgccat tttccagccg cgctcccacg agggtcacgg cggcggggag
     1021 aggtggagcc gcgagagctc ggccgggggc cccgcctggt ggtcgcggcc atgacagcgg
     1081 ctcgggacag gctccttttc cgcgcccctc ccgccggagg tgaggggaag atgtccatgt
     1141 cagggttcaa ggccaaaccg aagttactgg cctctatctt ccaggagaac caggagccac
     1201 agccgcggct cacgccccac cgcaacatta agattacaag tggacacctg agtcagcagg
     1261 acctggaatc ccagatgaga gagcttatct acacgactca gatcttgttg tcacccccat
     1321 tattgacaat ccaaaggtgc agaaagcact ctgacaaata atgaaacaac caccatcggt
     1381 taaatttgat gcaaaaatat tgcatctacc agcattttca gttccaattg ctaatccagc
     1441 atttgtggat agctgcaaac tgtgatattg ctgatgagcg ctttgacgcc acattccaca
     1501 ctaacgtgtt ggtgaattct tctgggcatt gccagtacct gcctccaggc atattcaaga
     1561 gttcctgcta catcgatgta cgctggtttc cctttgatgt gcagcactgc aaactgaagt
     1621 ttgggtcctg gtcttacgga ggctggtcct tggatctgca gatgcaggag gcagatatca
     1681 gtggctatat ccccaatgga gaatgggacc tagtgggaat ccccggcaag aggagtgaaa
     1741 ggttctatga gtgctgcaaa gagccctacc ctgatgtcac cttcacagtg accatgcgcc
     1801 gcaggacact ctactatggc ctcaacctgc tgatcccctg tgtgctcatc tccgccctcg
     1861 ccctgctggt gttcctgctt cctgcagatt ccggggagaa gatttccctg gggataacag
     1921 tcttactctc tcttaccgtc ttcatgctgc tcgtggctga gatcatgccc gcaacatccg
     1981 attcggtacc attgatagcc cagtacttcg ccagcaccat gatcatcgtg ggcctctcgg
     2041 tggtggtgac ggtgatcgtg ctgcagtacc accaccacga ccccgacggg ggcaagatgc
     2101 ccaagtggac cagagtcatc cttctgaact ggtgcgcgtg gttcctgcga atgaagaggc
     2161 ccggggagga caaggtgcgc ccggcctgcc agcacaagca gcggcgctgc agcctggcca
     2221 gtgtggagat gagcgccgtg gcgccgccgc ccgccagcaa cgggaacctg ctgtacatcg
     2281 gcttccgcgg cctggacggc gtgcactgtg tcccgacccc cgactctggg gtagtgtgtg
     2341 gccgcatggc ctgcgctggc gccatggtga gcaagggcga ggagctgttc accggggtgg
     2401 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2461 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2521 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2581 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2641 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2701 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2761 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2821 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2881 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2941 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagcttagc aaagacccca
     3001 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3061 gcatggacga gctgtacaag gcaggcgcct cccccacgca cgatgagcac ctcctgcacg
     3121 gcgggcaacc ccccgagggg gacccggact tggccaagat cctggaggag gtccgctaca
     3181 ttgccaaccg cttccgctgc caggacgaaa gcgaggcggt ctgcagcgag tggaagttcg
     3241 ccgcctgtgt ggtggaccgc ctgtgcctca tggccttctc ggtcttcacc atcatctgca
     3301 ccatcggcat cctgatgtcg gctcccaact tcgtggaggc cgtgtccaaa gactttgcgt
     3361 aaccacgcct ggttctgtac atgtggaaaa ctcacagatg ggcaaggaag ggcaattctg
     3421 cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc
     3481 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3541 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3601 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3661 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3721 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3781 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3841 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3901 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3961 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4021 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4081 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4141 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4201 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4321 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4381 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4441 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4501 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4561 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4621 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4681 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4741 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4801 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4861 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4921 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4981 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5041 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5101 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5161 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5221 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5281 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5341 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5401 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5461 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5521 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5701 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5761 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5821 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5881 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6661 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7741 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7801 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7861 catttccccg aaaagtgcca cctgacgtc
//
