LOCUS       sequence-313986-        7038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  AUTHORS   Nashmi R, Dickinson ME, McKinney S, Jareb M, Labarca C, Fraser SE, 
            Lester HA
  TITLE     Assembly of alpha4beta2 nicotinic acetylcholine receptors assessed 
            with functional fluorescently labeled subunits: effects of 
            localization, trafficking, and nicotine-induced upregulation in 
            clonal mammalian cells and in cultured midbrain neurons.
  JOURNAL   J Neurosci. 2003 Dec 17. 23(37):11554-67.
  PUBMED    14684858
REFERENCE   2  (bases 1 to 7038)
  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: "J Neurosci.
            2003 Dec 17. 23(37):11554-67."
COMMENT     Sequence Label: nAChR beta2 WT
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2605..2726
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2616..2635)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2670..2689
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2912..3367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2999..3018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3209..3230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3484..3841
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3484..3675
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3507..3527)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3692..3827
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3892..4686
                     /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(3946..3965)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4556..4575
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4750..4798
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4839..4858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4958..4980
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5018..5036)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5104..5208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5209..5277,5278..6069)
                     /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(5427..5446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6240..6828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6729..6748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcctcgaga
      961 attcgccctt ctccgcgcta tgcaggcggc atggcccggt gctccaactc tatggcgctg
     1021 ctgttcagct ttggcctcct ttggctgtgt tcaggggttt tgggtactga cacagaggag
     1081 cggctggtgg agcatctctt ggatccttcc cgctataaca agctgatccg tccagctact
     1141 aatggctctg agctggtgac tgtacagctc atggtatcat tggcacagct catcagtgtg
     1201 cacgagcggg agcagatcat gaccaccaac gtctggctga cccaggagtg ggaagattat
     1261 cgcctcacat ggaagcctga ggatttcgac aatatgaaga aagtccgact cccttctaag
     1321 cacatctggc tcccagatgt ggttctatac aacaatgctg acggcatgta cgaagtctcc
     1381 ttctattcca atgctgtggt ctcctatgat ggcagcatct tttggctacc gcctgccatc
     1441 tacaagagcg catgcaagat tgaggtgaag cacttcccat ttgaccagca gaactgcacc
     1501 atgaagttcc gctcctggac ctacgaccgc actgagattg acctggtgct caaaagcgat
     1561 gtggccagcc tggacgactt cacacctagt ggggagtggg acatcatcgc actgccaggc
     1621 cgacgcaacg agaacccaga cgactccacc tacgtggaca tcacctacga cttcatcatt
     1681 cgtcgcaaac cgctcttcta caccatcaac ctcatcatcc cctgcgtact catcacctcg
     1741 ctggccatcc tggtcttcta cctgccctca gactgtggtg aaaagatgac actttgtatt
     1801 tctgtgctgc tggcgctcac ggtgttcctg ctgctcatct ccaagattgt gcctcccacc
     1861 tccctcgacg taccgctggt gggaaagtac ctcatgttca ccatggtgct agtcaccttc
     1921 tccatcgtca ctagcgtgtg tgtgctcaat gtgcaccacc gttcgcctac cacgcacacc
     1981 atggcgccct gggtcaaggt ggtcttcctg gagaagctgc ccaccctcct cttcctgcag
     2041 cagccacgcc accgctgtgc acgtcaacgc ctgcgcttgc gaaggcgcca gcgggaacgt
     2101 gagggggcag gcacactatt cttccgcaaa ggtcctgcag ctgacccatg tacctgcttt
     2161 gtcaatcctg catcaatgca gggcttggct ggggctttcc aggctgagcc cgctgcagcc
     2221 ggtctggggc gctcgatggg cccatgcagc tgtggcctcc gggaagctgt ggacggtgta
     2281 cgcttcattg cggaccatat gcgaagtgaa gatgatgacc agagtgtgag ggaggattgg
     2341 aaatacgttg ccatggtgat cgaccgcctg ttcctctgga tctttgtctt tgtctgtgtc
     2401 tttgggacca ttggcatgtt cctgcagcct ctcttccaga actacactgc cacgaccttc
     2461 cttcactctg accactcagc tcccagctcc aagtgaggtc tctcatttgc agctcctcaa
     2521 gggcgaattc acgcgtggta cctctagagt cgacccgggc ggccgcttcc ctttagtgag
     2581 ggttaatgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     2641 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2701 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2761 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     2821 tccgataagg atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2881 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     2941 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3001 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3061 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3121 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3181 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3241 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3301 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3361 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3421 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3481 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3541 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3721 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3781 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3841 aaagcttgat tcttctgaca caacagtctc gaacttaagg ctagagccac catgattgaa
     3901 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3961 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4021 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4081 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4141 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4201 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4261 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4321 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4381 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4441 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4501 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4561 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4621 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4681 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4741 gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     4801 tcgatagcga taaggatccg cgtatggtgc actctcagta caatctgctc tgatgccgca
     4861 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4921 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4981 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     5041 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5101 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5161 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5221 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5281 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5341 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5401 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5461 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5521 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5581 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5641 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5701 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5761 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5821 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5881 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5941 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6001 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6061 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6121 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6181 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6241 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6301 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6361 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6421 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6481 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6541 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6601 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6661 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6721 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6781 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6841 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc
     6901 ttcaatattg gccattagcc atattattca ttggttatat agcataaatc aatattggct
     6961 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     7021 caatatgacc gccatgtt
//
