LOCUS       sequence-43766-e        7027 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 7027)
  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 7027)
  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:beta-2-365AAQA368 L349M
FEATURES             Location/Qualifiers
     source          1..7027
                     /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"
     polyA_signal    2731..2852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2742..2761)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2796..2815
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3038..3493
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3125..3144)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3335..3356
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3610..3967
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3610..3801
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3633..3653)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3818..3953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4018..4812
                     /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(4072..4091)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4682..4701
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4876..4924
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4965..4984)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5084..5106
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5144..5162)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5230..5334
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5335..5403,5404..6195)
                     /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(5553..5572)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6366..6954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6855..6874
                     /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 atgcagcagc cacgccaccg ctgtgcacgt caacgcctgc gcttgcgagc tgcccaggct
     2221 gaacgtgagg gggcaggcac actattcttc cgcgaaggtc ctgcagctga cccatgtacc
     2281 tgctttgtca atcctgcatc aatgcagggc ttggctgggg ctttccaggc tgagcccgct
     2341 gcagccggtc tggggcgctc gatgggccca tgcagctgtg gcctccggga agctgtggac
     2401 ggtgtacgct tcattgcgga ccatatgcga agtgaagatg atgaccagag tgtgagggag
     2461 gattggaaat acgttgccat ggtgatcgac cgcctgttcc tctggatctt tgtctttgtc
     2521 tgtgtctttg ggaccattgg catgttcctg cagcctctct tccagaacta cactgccacg
     2581 accttccttc actctgacca ctcagctccc agctccaagt gaggtctctc atttgcagct
     2641 cctcaagggc gaattcacgc gtggtacctc tagagtcgac ccgggcggcc gcttcccttt
     2701 agtgagggtt aatgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2761 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2821 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2881 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2941 gtaaaatccg ataaggatcg atccgggctg gcgtaatagc gaagaggccc gcaccgatcg
     3001 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     3061 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3121 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3181 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3241 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3301 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3361 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3421 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3481 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3541 gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt
     3601 aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3841 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3901 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3961 ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag agccaccatg
     4021 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4081 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4141 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4201 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4261 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4321 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4381 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4441 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4501 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4561 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4621 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4681 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4741 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4801 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4861 catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     4921 tgtgaatcga tagcgataag gatccgcgta tggtgcactc tcagtacaat ctgctctgat
     4981 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5041 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5101 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5161 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5221 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5281 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5341 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5401 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5461 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5521 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5581 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5641 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5701 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5761 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5821 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5881 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5941 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6001 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6061 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6121 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6181 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6241 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6301 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6361 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6421 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6481 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6541 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6601 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6661 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6721 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6781 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6841 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6901 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6961 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     7021 cagatct
//
