LOCUS       sequence-38130-e        7843 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 7843)
  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 7843)
  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: pmCherry-Sec24D
FEATURES             Location/Qualifiers
     source          1..7843
                     /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             26..733
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(171..189)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     439..458
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4047..4168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4084..4103)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4138..4157
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4175..4630)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4312..4333)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4524..4543
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4763..5120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4763..4954
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4786..4806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4971..5106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5155..5949
                     /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             5155..5949
                     /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(5209..5228)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5819..5838
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6137..6156)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6181..6228
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7046..7065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7320..7623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7624..7827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7778..7798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agatccgcta gcaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat
       61 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
      121 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      181 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
      241 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
      301 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
      361 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
      421 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
      481 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
      541 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
      601 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
      661 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
      721 cgagctgtac aagtccggac tcagatctcg agctcaagct tcgaattcca ccatgagtca
      781 acaaggttac gtggctacac ctccgtattc tcagcctcag cctggaatag gcctttctcc
      841 acctcattat gggcactatg gggatccgtc gcacacagca tctccaacag gtatgatgaa
      901 gccagcaggg cctttggggg ccaccgccac taggggaatg ttgcctccgg gtcccccacc
      961 tcctggaccc catcagtttg gtcagaatgg agctcatgcc actggtcacc ctccccaaag
     1021 atttccaggc cctccacctg tcaacaatgt ggcatcctca catgcaccat accaaccctc
     1081 tgcacaatct tcttatccag gtcctatatc cacttcatct gtcacccagc tgggcagcca
     1141 gctcagtgct atgcaaatca acagctatgg ttcaggcatg gctcctccaa gccagggacc
     1201 ccctggccct ctgtcagcca catcattgca gactcctcca cgacctccac agccttccat
     1261 tttgcagcct ggatctcaag ttcttccacc accacccacc acactcaatg gtcctggtgc
     1321 ctcacctttg cctctaccaa tgtacagacc agatgggctc tctgggcctc ctcctccaaa
     1381 tgcccagtac cagcccccac ctcttccagg ccagaccttg ggtgctggat atcctccgca
     1441 gcaggccaac tctggtcccc agatggcagg cgcacaactg tcttacccag gaggcttccc
     1501 tggaggtcct gcacagatgg ctggtccgcc acagccccag aagaagctgg atcctgactc
     1561 tatccctagc ccaatccagg tgattgagaa tgatagagcc agcagaggag gacaagttta
     1621 tgccaccaac accagaggcc agatccctcc cctggtcact acagattgca tgatacaaga
     1681 ccaaggaaat gccagtcctc gattcatccg ttgtacaaca tactgttttc catgcacgtc
     1741 agatatggct aagcaagctc agattccatt agctgctgtc atcaagccct ttgccaccat
     1801 tccttcaaat gagagtcccc tttacttggt aaatcacggc gagagtggac cagtcagatg
     1861 caacaggtgc aaggcctaca tgtgcccatt tatgcagttc atcgaaggag gaaggagata
     1921 tcagtgtgga ttttgcaact gtgtgaatga tgttccacca ttctatttcc aacatctgga
     1981 ccacattgga agaagactgg accactatga gaaaccagag ttatctctag gatcttatga
     2041 atatgttgcc actttggatt attgcagaaa gagtaagcct cccaacccac cagcctttat
     2101 cttcatgatt gatgtttcat atagtaacat aaagaatgga cttgtcaagc tcatatgtga
     2161 agaactgaag accatgctgg aaaaaattcc aaaggaagag caagaagaga cgtctgcaat
     2221 tcgagtgggt tttatcacat ataacaaagt tctccatttc tttaatgtga agagtaatct
     2281 ggcccagcct cagatgatgg tggtgactga tgttggagaa gtctttgttc ctttgttgga
     2341 tggtttcctt gtcaactatc aagaatccca atctgtgatt cataatttgt tggaccagat
     2401 tccagacatg tttgcagact ctaatgaaaa tgagactgtc tttgcttctg tcatccaggc
     2461 tggcatggaa gcactaaagg cagcagactg tcctgggaag ctgttcatct tccattcttc
     2521 cttgccaact gctgaagcac cagggaagct caaaaacaga gatgacaaaa aactggttaa
     2581 tacagacaaa gagaagatac ttttccagcc ccaaacaaat gtctatgact cattggccaa
     2641 ggactgcgtg gctcacggct gctctgtgac actcttcctc tttcctagtc agtatgtgga
     2701 cgtggcctcg ctggggctgg ttcctcagct cactggagga accctttaca aatacaacaa
     2761 tttccagatg cacttggata gacaacaatt tttgaacgac ctcagaaatg atattgaaaa
     2821 gaaaataggc tttgatgcta ttatgagggt tcgtaccagc acaggtttca gagccactga
     2881 tttctttggt ggaatcttga tgaacaacac caccgatgta gaaatggctg ccatcgattg
     2941 tgacaaggca gtgaccgtgg agttcaagca cgatgacaaa ctcagtgaag acagtggagc
     3001 cttaatccag tgtgctgtgc tttacacgac aatcagtggt caaagaagac ttcggattca
     3061 caatcttggc ttaaactgca gctctcagct agctgatctt tataagagct gtgagacaga
     3121 tgctcttatc aacttctttg ccaagtcagc ttttaaagca gttctccacc agcctttgaa
     3181 ggtcatccgg gaaattctag ttaatcagac tgcccatatg ttggcatgtt accggaagaa
     3241 ttgtgcaagt ccttctgcag caagccagct tattctacca gattccatga aagtattgcc
     3301 agtgtacatg aattgcttgt tgaaaaactg tgtactactc agcagaccag agatctcaac
     3361 tgatgaacga gcataccaga gacagctggt catgaccatg ggtgtggctg actctcagct
     3421 tttcttctac ccacaacttc tgcccataca cacgttagat gtcaagagta caatgttacc
     3481 tgctgccgtt cgttgctctg agtcccgtct ttcagaagaa ggaatattct tactggctaa
     3541 tggtctacac atgttcctgt ggttgggagt aagcagccca ccagaactga tccaaggaat
     3601 atttaatgtg ccatcttttg cacatatcaa cacagatatg acattgctgc ctgaagtggg
     3661 aaacccatac tctcaacaac tcagaatgat aatgggtatt atccaacaaa agaggccata
     3721 ttcaatgaag ctcacaattg taaagcagcg agaacaacca gaaatggttt tccgacagtt
     3781 cctggtagaa gacaaaggac tttacggagg ctcttcttat gtggatttcc tttgttgtgt
     3841 tcacaaggag atctgtcagc tgcttaatta attgaaactt ctctgtgatt gatgttgcat
     3901 ttccaaggag ataatctcct tcttggtgcc taattttcta gatcataatc agccatacca
     3961 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4021 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4081 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4141 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4201 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4261 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4321 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4381 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4441 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4501 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4561 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4621 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4801 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4861 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4921 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4981 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5041 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5101 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5161 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5221 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5281 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5341 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5401 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5461 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5521 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5581 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5641 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5701 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5761 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5821 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5881 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5941 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6001 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6061 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6121 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6181 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6241 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6301 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6361 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     7261 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7321 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7381 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7441 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7501 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7561 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7621 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     7681 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     7741 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     7801 cggtgggagg tctatataag cagagctggt ttagtgaacc gtc
//
