LOCUS       sequence-62577-e        8150 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 8150)
  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 8150)
  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-mcherry
FEATURES             Location/Qualifiers
     source          1..8150
                     /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             2384..3091
                     /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(2529..2547)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2797..2816
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3854..3975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3865..3884)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3919..3938
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4161..4616
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4248..4267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4458..4479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4733..5090
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4733..4924
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4756..4776)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4941..5076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5141..5935
                     /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(5195..5214)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5805..5824
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5999..6047
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6088..6107)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6207..6229
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6267..6285)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6353..6457
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6458..6526,6527..7318)
                     /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(6676..6695)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7489..8077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7978..7997
                     /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 cagtcgagca acagcctaca tgcatggtga gcaagggcga
     2401 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     2461 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     2521 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     2581 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     2641 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     2701 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     2761 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     2821 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     2881 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     2941 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3001 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3061 ctccaccggc ggcatggacg agctgtacaa gaggtcaccc tcccacaagg tccctgactt
     3121 gaagacatca gaggttgaga aggccagtcc ctgtccatca cctggctctt gtcacccacc
     3181 caatagcagt ggggccccag tgctcatcaa agccaggtcc ctgagcgtcc agcatgtgcc
     3241 cagctcccag gaagcagccg agggcagcat ccgctgccgg tctcggagta tccagtactg
     3301 tgtttcccaa gatggagctg cttccctgac tgagagcaag cccactggct ccccagcctc
     3361 cctgaagacc cgtccatccc agcttccagt gtcagaccag acctctccat gcaaatgcac
     3421 atgcaaggaa ccatctcctg tgtcccccat cactgtgctc aaggctggag gcaccaaagc
     3481 acctccccaa cacctgcccc tgtcaccagc cctgacacgg gcagtagaag gcgtccagta
     3541 cattgcagac cacctcaagg cagaagacac agacttctcg gtgaaggagg actggaaata
     3601 cgtggccatg gtcattgacc gaatcttcct ctggatgttc atcattgtct gccttctggg
     3661 cactgtggga ctcttcctgc ctccatggtt ggctggtatg atctagggaa tagcggcacc
     3721 tagctcccag gtctctacag ggccatgcga ctcgtcagtc cacccacaag ggcgaattca
     3781 cgcgtggtac ctctagagtc gacccgggcg gccgcttccc tttagtgagg gttaatgctt
     3841 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3901 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3961 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     4021 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat ccgataagga
     4081 tcgatccggg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4141 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4381 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4441 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4501 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4561 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     4621 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     4681 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     4741 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4861 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4921 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4981 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5041 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt
     5101 cttctgacac aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt
     5161 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     5221 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     5281 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     5341 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     5401 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5461 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5521 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5581 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5641 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5701 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5761 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5821 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5881 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5941 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     6001 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     6061 aaggatccgc gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6121 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6181 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6241 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6301 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6361 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6421 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6481 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6541 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6601 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6661 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6721 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6781 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6841 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6901 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6961 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7021 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7081 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7141 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7201 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7261 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7321 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7381 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7441 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7561 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7621 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7921 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7981 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8041 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8101 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct
//
