LOCUS       sequence-38143-e        7852 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 7852)
  AUTHORS   Richards CI, Srinivasan R, Xiao C, Mackey ED, Miwa JM, Lester HA
  TITLE     Trafficking of {alpha}4* Nicotinic Receptors Revealed by 
            Superecliptic Phluorin: EFFECTS OF A {beta}4 AMYOTROPHIC LATERAL 
            SCLEROSIS-ASSOCIATED MUTATION AND CHRONIC EXPOSURE TO NICOTINE.
  JOURNAL   J Biol Chem. 2011 Sep 9;286(36):31241-9. Epub 2011 Jul 18.
  PUBMED    21768117
REFERENCE   2  (bases 1 to 7852)
  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 Biol 
            Chem."; date: "2011-09-9"; volume: "286(36)"; pages: "31241-9. Epub 
            2011 Jul 18"
COMMENT     Sequence Label: pEGFP-Sec24D
FEATURES             Location/Qualifiers
     source          1..7852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      20..29
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(26..28,29..31,32..742)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(26..28,29..31,32..742)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(71..92)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(332..351)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     679..700
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4056..4177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4093..4112)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4147..4166
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4184..4639)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4321..4342)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4533..4552
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4666..4770
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4772..5129
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4772..4963
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4795..4815)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4980..5115
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5164..5958
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5164..5958
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5218..5237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5828..5847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6146..6165)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6190..6237
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6566..7154
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7055..7074
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7329..7632
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7633..7836
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7787..7807
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agatccgcta gcaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
       61 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      121 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      181 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
      241 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      301 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      361 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      421 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
      481 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
      541 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
      601 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
      661 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
      721 cggcatggac gagctgtaca agtccggact cagatctcga gctcaagctt cgaattccac
      781 catgagtcaa caaggttacg tggctacacc tccgtattct cagcctcagc ctggaatagg
      841 cctttctcca cctcattatg ggcactatgg ggatccgtcg cacacagcat ctccaacagg
      901 tatgatgaag ccagcagggc ctttgggggc caccgccact aggggaatgt tgcctccggg
      961 tcccccacct cctggacccc atcagtttgg tcagaatgga gctcatgcca ctggtcaccc
     1021 tccccaaaga tttccaggcc ctccacctgt caacaatgtg gcatcctcac atgcaccata
     1081 ccaaccctct gcacaatctt cttatccagg tcctatatcc acttcatctg tcacccagct
     1141 gggcagccag ctcagtgcta tgcaaatcaa cagctatggt tcaggcatgg ctcctccaag
     1201 ccagggaccc cctggccctc tgtcagccac atcattgcag actcctccac gacctccaca
     1261 gccttccatt ttgcagcctg gatctcaagt tcttccacca ccacccacca cactcaatgg
     1321 tcctggtgcc tcacctttgc ctctaccaat gtacagacca gatgggctct ctgggcctcc
     1381 tcctccaaat gcccagtacc agcccccacc tcttccaggc cagaccttgg gtgctggata
     1441 tcctccgcag caggccaact ctggtcccca gatggcaggc gcacaactgt cttacccagg
     1501 aggcttccct ggaggtcctg cacagatggc tggtccgcca cagccccaga agaagctgga
     1561 tcctgactct atccctagcc caatccaggt gattgagaat gatagagcca gcagaggagg
     1621 acaagtttat gccaccaaca ccagaggcca gatccctccc ctggtcacta cagattgcat
     1681 gatacaagac caaggaaatg ccagtcctcg attcatccgt tgtacaacat actgttttcc
     1741 atgcacgtca gatatggcta agcaagctca gattccatta gctgctgtca tcaagccctt
     1801 tgccaccatt ccttcaaatg agagtcccct ttacttggta aatcacggcg agagtggacc
     1861 agtcagatgc aacaggtgca aggcctacat gtgcccattt atgcagttca tcgaaggagg
     1921 aaggagatat cagtgtggat tttgcaactg tgtgaatgat gttccaccat tctatttcca
     1981 acatctggac cacattggaa gaagactgga ccactatgag aaaccagagt tatctctagg
     2041 atcttatgaa tatgttgcca ctttggatta ttgcagaaag agtaagcctc ccaacccacc
     2101 agcctttatc ttcatgattg atgtttcata tagtaacata aagaatggac ttgtcaagct
     2161 catatgtgaa gaactgaaga ccatgctgga aaaaattcca aaggaagagc aagaagagac
     2221 gtctgcaatt cgagtgggtt ttatcacata taacaaagtt ctccatttct ttaatgtgaa
     2281 gagtaatctg gcccagcctc agatgatggt ggtgactgat gttggagaag tctttgttcc
     2341 tttgttggat ggtttccttg tcaactatca agaatcccaa tctgtgattc ataatttgtt
     2401 ggaccagatt ccagacatgt ttgcagactc taatgaaaat gagactgtct ttgcttctgt
     2461 catccaggct ggcatggaag cactaaaggc agcagactgt cctgggaagc tgttcatctt
     2521 ccattcttcc ttgccaactg ctgaagcacc agggaagctc aaaaacagag atgacaaaaa
     2581 actggttaat acagacaaag agaagatact tttccagccc caaacaaatg tctatgactc
     2641 attggccaag gactgcgtgg ctcacggctg ctctgtgaca ctcttcctct ttcctagtca
     2701 gtatgtggac gtggcctcgc tggggctggt tcctcagctc actggaggaa ccctttacaa
     2761 atacaacaat ttccagatgc acttggatag acaacaattt ttgaacgacc tcagaaatga
     2821 tattgaaaag aaaataggct ttgatgctat tatgagggtt cgtaccagca caggtttcag
     2881 agccactgat ttctttggtg gaatcttgat gaacaacacc accgatgtag aaatggctgc
     2941 catcgattgt gacaaggcag tgaccgtgga gttcaagcac gatgacaaac tcagtgaaga
     3001 cagtggagcc ttaatccagt gtgctgtgct ttacacgaca atcagtggtc aaagaagact
     3061 tcggattcac aatcttggct taaactgcag ctctcagcta gctgatcttt ataagagctg
     3121 tgagacagat gctcttatca acttctttgc caagtcagct tttaaagcag ttctccacca
     3181 gcctttgaag gtcatccggg aaattctagt taatcagact gcccatatgt tggcatgtta
     3241 ccggaagaat tgtgcaagtc cttctgcagc aagccagctt attctaccag attccatgaa
     3301 agtattgcca gtgtacatga attgcttgtt gaaaaactgt gtactactca gcagaccaga
     3361 gatctcaact gatgaacgag cataccagag acagctggtc atgaccatgg gtgtggctga
     3421 ctctcagctt ttcttctacc cacaacttct gcccatacac acgttagatg tcaagagtac
     3481 aatgttacct gctgccgttc gttgctctga gtcccgtctt tcagaagaag gaatattctt
     3541 actggctaat ggtctacaca tgttcctgtg gttgggagta agcagcccac cagaactgat
     3601 ccaaggaata tttaatgtgc catcttttgc acatatcaac acagatatga cattgctgcc
     3661 tgaagtggga aacccatact ctcaacaact cagaatgata atgggtatta tccaacaaaa
     3721 gaggccatat tcaatgaagc tcacaattgt aaagcagcga gaacaaccag aaatggtttt
     3781 ccgacagttc ctggtagaag acaaaggact ttacggaggc tcttcttatg tggatttcct
     3841 ttgttgtgtt cacaaggaga tctgtcagct gcttaattaa ttgaaacttc tctgtgattg
     3901 atgttgcatt tccaaggaga taatctcctt cttggtgcct aattttctag atcataatca
     3961 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4021 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4081 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4141 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     4201 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4261 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4321 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4381 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4441 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4501 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4561 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4621 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4741 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4801 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4861 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4921 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4981 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5041 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5101 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5161 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5221 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5281 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5341 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5401 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5461 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5521 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5581 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5641 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5701 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5761 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5821 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5881 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5941 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6001 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6061 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6121 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6181 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6241 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6301 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6361 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6421 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6481 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6541 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6601 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6661 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6721 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6781 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6841 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6901 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6961 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7021 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7081 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7141 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7201 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7261 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7321 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7381 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7441 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7501 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7561 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     7621 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     7681 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     7741 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     7801 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tc
//
