LOCUS       sequence-62591-e        8195 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 8195)
  AUTHORS   Srinivasan R, Pantoja R, Moss FJ, Mackey ED, Son CD, Miwa J, Lester 
            HA
  TITLE     Nicotine up-regulates alpha4beta2 nicotinic receptors and ER exit 
            sites via stoichiometry-dependent chaperoning.
  JOURNAL   J Gen Physiol. 2011 Jan;137(1):59-79.
  PUBMED    21187334
REFERENCE   2  (bases 1 to 8195)
  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 Gen 
            Physiol."; date: "2011-01"; volume: "137(1)"; pages: "59-79"
COMMENT     Sequence Label: pCI-neo:alpha4-eGFP
FEATURES             Location/Qualifiers
     source          1..8195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2393..2419
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2420..2422,2423..2425,2426..3136)
                     /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(2465..2486)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2726..2745)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3073..3094
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3899..4020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3910..3929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3964..3983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4206..4661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4293..4312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4503..4524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4778..5135
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4778..4969
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4801..4821)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4986..5121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5186..5980
                     /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(5240..5259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5850..5869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6044..6092
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6133..6152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6252..6274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6312..6330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6398..6502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6503..6571,6572..7363)
                     /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(6721..6740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7534..8122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8023..8042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt cggcttccat ggagatcggg ggctccgggg cgccgccgcc
     1141 gctgctgctc ctgccgctcc tgctgctctt agggaccggc ctcttgcctg ctagcagcca
     1201 catagagacc cgggcccatg cggaggagcg gctcctgaag agactcttct ctggctacaa
     1261 caagtggtct cggccagtag ccaatatctc agatgtggtc cttgtccgct ttggcttgtc
     1321 gattgctcag ctcattgatg tggatgagaa aaaccagatg atgacgacca acgtgtgggt
     1381 gaagcaggag tggcatgact acaaactgcg ctgggaccct ggtgactacg agaatgtcac
     1441 ctccatccgc atcccatctg aactcatctg gaggcctgac atcgtcctct acaacaacgc
     1501 ggacggggac tttgcagtca cccacctaac caaagcccac ctgttctatg atgggcgtgt
     1561 acagtggaca cccccggcca tctataagag ctcctgcagc atcgacgtca ccttcttccc
     1621 cttcgaccag cagaactgta ccatgaagtt tgggtcctgg acctacgaca aggccaagat
     1681 tgacttggtg agcatgcaca gccgtgtgga ccaactggac ttctgggaaa gtggggagtg
     1741 ggtcattgtg gatgccgtgg gcacctacaa caccaggaag tatgaatgct gtgccgagat
     1801 ctatcctgac atcacctacg ccttcatcat ccgccgactg ccactgttct acaccatcaa
     1861 ccttatcatc ccgtgcctgc tcatctcctg cctcaccgtg ctggtcttct atctgccctc
     1921 ggagtgcggc gagaaggtca cgctgtgcat ctcggtgctg ctttctctca ccgtcttcct
     1981 gctgctcatc accgaaatca tcccgtccac ctcgctggtc atcccgctca tcggcgagta
     2041 cctgctcttc accatgatct tcgtcaccct ctccattgtc atcacggtct tcgtgctcaa
     2101 tgtacaccac cgctcaccac gcacacacac catgcccgcc tgggtgcgca gagtcttcct
     2161 ggacattgtg ccccgtctcc tcttcatgaa gcgcccatct gtggtcaaag acaactgccg
     2221 gagacttatc gaatccatgc acaagatggc caacgcccct cgtttctggc cagagcctga
     2281 gagtgagccc ggcatcttgg gtgacatctg caaccaaggc ctgtcacctg ccccaacttt
     2341 ctgcaaccgc atggacacag cagtcgagac ccagcctaca tgcaggtcac cctatcctta
     2401 tgacgtccca gactacgcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2461 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2521 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2581 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2641 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2701 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2761 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2821 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2881 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2941 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3001 gctgctgccc gacaaccact acctgagcac ccagtccaag cttagcaaag accccaacga
     3061 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3121 ggacgagctg tacaagaggt caccctccca caaggtccct gacttgaaga catcagaggt
     3181 tgagaaggcc agtccctgtc catcacctgg ctcttgtcac ccacccaata gcagtggggc
     3241 cccagtgctc atcaaagcca ggtccctgag cgtccagcat gtgcccagct cccaggaagc
     3301 agccgagggc agcatccgct gccggtctcg gagtatccag tactgtgttt cccaagatgg
     3361 agctgcttcc ctgactgaga gcaagcccac tggctcccca gcctccctga agacccgtcc
     3421 atcccagctt ccagtgtcag accagacctc tccatgcaaa tgcacatgca aggaaccatc
     3481 tcctgtgtcc cccatcactg tgctcaaggc tggaggcacc aaagcacctc cccaacacct
     3541 gcccctgtca ccagccctga cacgggcagt agaaggcgtc cagtacattg cagaccacct
     3601 caaggcagaa gacacagact tctcggtgaa ggaggactgg aaatacgtgg ccatggtcat
     3661 tgaccgaatc ttcctctgga tgttcatcat tgtctgcctt ctgggcactg tgggactctt
     3721 cctgcctcca tggttggctg gtatgatcta gggaatagcg gcacctagct cccaggtctc
     3781 tacagggcca tgcgactcgt cagtccaccc acaagggcga attcacgcgt ggtacctcta
     3841 gagtcgaccc gggcggccgc ttccctttag tgagggttaa tgcttcgagc agacatgata
     3901 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3961 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4021 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     4081 taaagcaagt aaaacctcta caaatgtggt aaaatccgat aaggatcgat ccgggctggc
     4141 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4201 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4261 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4321 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4381 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4441 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4501 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4561 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4621 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     4681 cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg
     4741 aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt
     4801 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4861 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4921 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4981 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5041 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5101 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag
     5161 tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc
     5221 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5281 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5341 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5401 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5461 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5521 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5581 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5641 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5701 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5761 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5821 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5881 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5941 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6001 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     6061 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg
     6121 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6181 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6241 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6301 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6361 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7561 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7621 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7681 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7741 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7801 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7861 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7921 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7981 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8041 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8101 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8161 ttgctggcct tttgctcaca tggctcgaca gatct
//
