LOCUS       sequence-106765-        7887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human duplicated Alpha7-2 bp deletion 
            (CHRFAM7A-∆2bp) with YFP fused into M3-M4 loop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7887)
  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 7887)
  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-CHRFAM7ADelta2bp-YFP
FEATURES             Location/Qualifiers
     source          1..7887
                     /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(2362..2364,2365..2367,2368..3078)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2407..2428)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2668..2687)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3015..3036
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3481..3498)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3487..3711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3757..4185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3844..3863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4054..4075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4194..4214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4199..4528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4379..4514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4441..4460
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4595..5389
                     /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(4649..4668)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5259..5278
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5563..5684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5600..5619)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5654..5673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5733..5749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5733..5749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5746..5768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5757..5773
                     /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(5781..5787,5788..5805,5806..5811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5826..5847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5964..5981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6135..6720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6215..6234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6891..7682,7683..7751))
                     /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     7514..7533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7752..7856)
                     /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 aaacaagttt
     1621 gggtcctggt cttacggagg ctggtccttg gatctgcaga tgcaggaggc agatatcagt
     1681 ggctatatcc ccaatggaga atgggaccta gtgggaatcc ccggcaagag gagtgaaagg
     1741 ttctatgagt gctgcaaaga gccctaccct gatgtcacct tcacagtgac catgcgccgc
     1801 aggacactct actatggcct caacctgctg atcccctgtg tgctcatctc cgccctcgcc
     1861 ctgctggtgt tcctgcttcc tgcagattcc ggggagaaga tttccctggg gataacagtc
     1921 ttactctctc ttaccgtctt catgctgctc gtggctgaga tcatgcccgc aacatccgat
     1981 tcggtaccat tgatagccca gtacttcgcc agcaccatga tcatcgtggg cctctcggtg
     2041 gtggtgacgg tgatcgtgct gcagtaccac caccacgacc ccgacggggg caagatgccc
     2101 aagtggacca gagtcatcct tctgaactgg tgcgcgtggt tcctgcgaat gaagaggccc
     2161 ggggaggaca aggtgcgccc ggcctgccag cacaagcagc ggcgctgcag cctggccagt
     2221 gtggagatga gcgccgtggc gccgccgccc gccagcaacg ggaacctgct gtacatcggc
     2281 ttccgcggcc tggacggcgt gcactgtgtc ccgacccccg actctggggt agtgtgtggc
     2341 cgcatggcct gcgctggcgc catggtgagc aagggcgagg agctgttcac cggggtggtg
     2401 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2461 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2521 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     2581 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2641 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2701 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2761 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2821 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2881 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2941 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     3001 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3061 atggacgagc tgtacaaggc aggcgcctcc cccacgcacg atgagcacct cctgcacggc
     3121 gggcaacccc ccgaggggga cccggacttg gccaagatcc tggaggaggt ccgctacatt
     3181 gccaaccgct tccgctgcca ggacgaaagc gaggcggtct gcagcgagtg gaagttcgcc
     3241 gcctgtgtgg tggaccgcct gtgcctcatg gccttctcgg tcttcaccat catctgcacc
     3301 atcggcatcc tgatgtcggc tcccaacttc gtggaggccg tgtccaaaga ctttgcgtaa
     3361 ccacgcctgg ttctgtacat gtggaaaact cacagatggg caaggaaggg caattctgca
     3421 gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag
     3481 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3541 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3601 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3661 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3721 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     3781 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3841 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3901 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3961 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4021 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4081 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4141 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4201 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4261 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4321 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4381 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4441 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     4501 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     4561 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4621 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4681 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4741 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4801 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4861 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4921 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4981 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5041 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5101 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5161 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5221 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5281 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5341 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5401 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5461 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5521 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5581 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5641 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5701 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5821 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5881 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5941 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6001 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6061 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6121 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6181 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6241 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6301 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6361 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6421 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6481 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6541 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6601 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6661 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6781 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7021 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7861 tttccccgaa aagtgccacc tgacgtc
//
