LOCUS       sequence-62596-e        7780 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 7780)
  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 7780)
  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-neoBeta2mcherry
FEATURES             Location/Qualifiers
     source          1..7780
                     /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             2263..2292
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2293..3000
                     /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(2438..2456)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2706..2725
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3484..3605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3495..3514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3549..3568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3791..4246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3878..3897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4088..4109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4363..4720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4363..4554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4386..4406)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4571..4706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4771..5565
                     /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(4825..4844)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5435..5454
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5629..5677
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5718..5737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5837..5859
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5897..5915)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5983..6087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6088..6156,6157..6948)
                     /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(6306..6325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7119..7707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7608..7627
                     /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 ccgctatgta agcggaatgg cccggtgctc caactctatg
     1141 gggctgctgt tcagctttgg cctcctttgg ctgtgttcag gggttttggg tactgacaca
     1201 cacgagcggc tggtggagca tctcttggat ccttcccgct ataacaagct gatccgacca
     1261 gctactaatg gctctgagct ggtgactgta cagctcatgg tatcattggc acagctcatc
     1321 agtgtgcacg agcgggagca gatcatgacc accaacgtct ggctgaccca ggagtgggaa
     1381 gattatcgcc tcacatggaa gcctgaggat ttcgacaata tgaagaaagt ccgactccct
     1441 tctaagcaca tctggctccc agatgtggtt ctatacaaca atgctgacgg catgtacgaa
     1501 gtctccttct attccaatgc tgtggtctcc tatgatggca gcatcttttg gctaccgcct
     1561 gccatctaca agatcgcatg caagattgag gtgaagcact tcccatttga ccakcagaac
     1621 tgcaccatga agttccgctc ctggacctac gaccgcactg agattgacct grtgctcwaa
     1681 agckatgtgg ccagcctgga cgacttcaca cctagtgggg agtgggacat catcgcactg
     1741 tcaggccgac gcaacgagaa cccagacgac tccacctacg tggacatcac ctacgacttc
     1801 atcattcgtc gmaaaccgmt cttctacacc atcaacctca tcatcccctg cgtactcatc
     1861 acctcgctgg scatcctggt cttctacctg ccctcagact gtggtgaaaa gatgacactt
     1921 tgtatttctg tgctgctggc gctcacggtg ttcctgctgc tcatctccaa gattgtgcct
     1981 cccacctccc tcgacgtacc gctggtggga aagtacctca tgttcaccat ggtgctagtc
     2041 accttctcca tcgtcactag cgtgtgtgtg ctcaatgtgc accaccgttc gcctaccacg
     2101 cacaccatgg cgccctgggt caaggtggtc ttcctggaga agctgcccac cctcctcttc
     2161 ctgcagcagc cacgccaccg ctgtgcacgt caacgcctgc gcttgcgaag gcgccagcgg
     2221 gaacgtgagg gggcaggcac actattcttc cgcaaaggtc ctgagcagaa gctgatctca
     2281 gaggaggatc tgatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2341 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     2401 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     2461 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     2521 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2581 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2641 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     2701 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     2761 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2821 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2881 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     2941 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3001 tcctggtcct gtcctgcagc tgacccatgt acctgctttg tcaatcctgc atcaatgcag
     3061 ggcttggctg gggctttcca ggctgagccc gctgcagccg gtctggggcg ctcgatgggc
     3121 ccatgcagct gtggcctccg ggaagctgtg gacggtgtac gcttcattgc ggaccatatg
     3181 cgaagtgaag atgatgacca gagtgtgagg gaggattgga aatacgttgc catggtgatc
     3241 gaccgcctgt tcctctggat ctttgtcttt gtctgtgtct ttgggaccat tggcatgttc
     3301 ctgcagcctc tcttccagaa ctacactgcc acgaccttcc ttcactctga ccactcagct
     3361 cccagctcca agtgaggtct ctcatttgca gctcctcaag ggcgaattca cgcgtggtac
     3421 ctctagagtc gacccgggcg gccgcttccc tttagtgagg gttaatgctt cgagcagaca
     3481 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3541 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3601 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3661 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat ccgataagga tcgatccggg
     3721 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3781 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3841 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3901 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3961 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4021 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4081 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4141 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4201 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     4261 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     4321 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     4381 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4441 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4501 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4561 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4621 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4681 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac
     4741 aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt
     4801 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4861 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4921 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4981 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5041 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5101 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5161 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5221 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5281 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5341 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5401 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5461 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5521 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5581 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct
     5641 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc
     5701 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5761 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5821 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5881 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5941 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6001 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6061 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6121 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6181 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6241 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6301 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6361 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6421 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6481 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6541 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6601 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6661 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6721 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6781 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6841 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6901 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6961 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7021 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7081 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7141 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7201 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7261 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7321 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7381 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7441 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7501 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7561 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7621 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7681 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7741 gccttttgct ggccttttgc tcacatggct cgacagatct
//
