LOCUS       sequence-67919-e        8811 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 8811)
  AUTHORS   Chicka MC, Strehler EE
  TITLE     Alternative splicing of the first intracellular loop of plasma 
            membrane Ca2+-ATPase isoform 2 alters its membrane targeting.
  JOURNAL   J Biol Chem. 2003 May 16;278(20):18464-70. Epub 2003 Mar 6.
  PUBMED    12624087
REFERENCE   2  (bases 1 to 8811)
  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: "2003-05-16"; volume: "278(20)"; pages: "18464-70. 
            Epub 2003 Mar 6"
COMMENT     Sequence Label: pMM2-hPMCA2z/b
FEATURES             Location/Qualifiers
     source          1..8811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(7..336)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      21..156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     321..341
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          805..1034
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1017..1036
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1605..1616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2031..2040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4859..5422
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    5610..5744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5647..5666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5701..5720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6417..6434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6588..7176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6668..6687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7347..8138,8139..8207))
                     /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     7970..7989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8208..8312)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8380..8398
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8436..8458)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8558..8577
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8771..8793
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8785..8802
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8786..8802
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
       61 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
      121 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
      181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      241 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      301 ggagcctggg gactttccac accctaactg acacacattc cacaggatcc ggtcgcgcga
      361 atttcgagcg gtgttccgcg gtcctcctcg tatagaaact cggaccactc tgagacgaag
      421 gctcgcgtcc aggccagcac gaaggaggct aagtgggagg ggtagcggtc gttgtccact
      481 agggggtcca ctcgctccag ggtgtgaaga cacatgtcgc cctcttcggc atcaaggaag
      541 gtgattggtt tataggtgta ggccacgtga ccgggtgttc ctgaaggggg gctataaaag
      601 ggggtggggg cgcgttcgtc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt
      661 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc
      721 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga
      781 tcggaaaacc tctcgactgt tggggtgagt actccctctc aaaagcgggc atgacttctg
      841 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga
      901 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct
      961 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc
     1021 ctttctctcc acaggtgtcc actcccaggt ccaactgcag gtcgacacgc gtacgtaagc
     1081 ttggatcctc tagagcggcc gccatgggtg acatgaccaa cagcgacttt tactccaaaa
     1141 accaaagaaa tgagtcgagc catgggggcg agttcgggtg cacaatggag gagctccgct
     1201 ccctcatgga gctgcggggc actgaggctg tggtcaagat caaggagact tatggggaca
     1261 ccgaagccat ctgccggcgc ctcaaaacct cacctgttga aggtttgccg ggcaccgctc
     1321 cagacctgga aaagagaaag caaatttttg ggcaaaactt tatacctcca aagaagccaa
     1381 aaaccttcct gcagctcgtg tgggaggcgc tgcaggacgt gacgctcatc atcctggaga
     1441 ttgccgccat catctccctg gggctgtcct tctaccaccc gcccggcgag ggcaacgaag
     1501 gatgtgcgac ggcccagggt ggggcagagg atgaaggaga ggcagaggca ggttggatcg
     1561 agggggccgc cattctcctc tcagttatct gtgtggtcct ggtcacggcc ttcaatgact
     1621 ggagcaaaga gaaacagttc cggggcctgc agagccgcat cgagcaggaa cagaaattta
     1681 ccgtggtccg ggctggccag gtggtgcaga tccctgtggc tgagatcgtg gttggggaca
     1741 tagcccaggt caaatatggt gacctcctcc ctgccgacgg cctcttcatc cagggcaatg
     1801 acctcaagat tgatgaaagc tccctaactg gagagtctga ccaggtgcgc aagtccgtgg
     1861 acaaggaccc catgctgctg tcaggaaccc acgtgatgga gggctcagga cggatgttgg
     1921 tgactgctgt gggtgtgaac tctcagactg gcatcatctt taccctcctg ggggctggtg
     1981 gtgaagagga agagaagaaa gacaaaaaag ccaaacaaca ggacggggca gccgccatgg
     2041 agatgcagcc cctcaagagt gccgagggcg gcgacgctga cgacaggaag aaggccagca
     2101 tgcacaagaa ggagaagtcc gtgctgcagg gcaagctcac caagctggct gtgcagatcg
     2161 ggaaggcggg cttggtgatg tcagccatca cggtgatcat cctggtgctc tacttcactg
     2221 tggacacctt cgtggtcaac aagaagccgt ggctgcctga gtgcacgccc gtctacgtgc
     2281 agtactttgt caagttcttc atcattggcg tgacggtgct ggtggtcgcc gtgcccgagg
     2341 ggctccctct ggccgtcacc atctcgttgg cctattcggt gaagaaaatg atgaaggaca
     2401 acaacctggt acgccacctg gatgcctgtg agaccatggg caatgccaca gccatctgct
     2461 cagacaagac aggcacgctg accaccaatc gcatgacagt ggtacaggcc tatgtcggcg
     2521 acgtccacta taaagagatc cccgacccca gctccatcaa caccaagacc atggagctgc
     2581 tgatcaatgc catcgccatc aacagcgcct acaccaccaa gattctgccc ccagagaagg
     2641 agggcgccct gcctcggcag gtgggcaaca agacggagtg cggcctgctg ggcttcgtgc
     2701 tggacctgaa gcaggactac gagcccgtgc gcagccagat gccagaggag aagttgtaca
     2761 aagtgtacac cttcaactcc gtgcgcaagt ccatgagcac tgtcatcaag ctgcccgacg
     2821 agagcttccg catgtacagc aagggggctt ctgagatcgt gctcaagaag tgctgcaaaa
     2881 tcctcaatgg ggcgggagag cctcgtgtct tccggccccg cgaccgggac gagatggtaa
     2941 agaaggtgat tgagcccatg gcttgcgatg ggctccgcac tatctgcgtg gcctaccgcg
     3001 acttccccag cagcccggag ccggactggg acaatgagaa tgacatcctc aacgaactca
     3061 cctgcatctg cgtggtgggc atcgaggacc cggtgcggcc agaggtccca gaagccatcc
     3121 gcaagtgcca gcgggcaggc atcacggtcc gcatggtcac tggcgacaat atcaacacgg
     3181 ctcgggccat cgccatcaag tgtggcatca tccatcctgg ggaggacttt ctgtgcctcg
     3241 agggcaagga gttcaacagg aggatccgca acgagaaggg ggagattgag caggagcgaa
     3301 ttgacaagat ctggccaaag ctgcgggtgc tggctcgctc ctccccaacg gacaagcata
     3361 ccctggttaa aggcatcatc gacagcacac acactgagca gcggcaggtg gtggccgtga
     3421 cgggggacgg gaccaacgac gggcctgcac tcaagaaggc cgacgtgggc ttcgccatgg
     3481 gcatcgcagg cactgacgtg gccaaggagg cctcagacat catcctgaca gacgacaatt
     3541 tcagcagcat cgtcaaggca gtgatgtggg gccgcaacgt ctatgacagc atctccaaat
     3601 tcttgcagtt ccagctcacc gtcaacgtgg tggccgtgat tgtggccttc acaggcgcct
     3661 gcatcacgca ggactcccct ctgaaggccg tgcagatgct ctgggtgaac ctcatcatgg
     3721 acacgtttgc ctcgctggca ctggccactg agccgcccac ggagaccctg ctgctgagga
     3781 agccgtacgg ccgcaacaag ccgctcatct ccaggaccat gatgaagaac atcctgggcc
     3841 atgctgtcta ccagcttgcc ctcatcttca ccctgctctt tgttggcgag aagatgttcc
     3901 agatcgacag cgggaggaac gcgcccctgc attcgccacc ctcagaacat tacaccatca
     3961 tcttcaacac cttcgtcatg atgcagctct tcaacgagat caacgcccgc aagatccacg
     4021 gcgagcgcaa tgtctttgac ggcatcttcc ggaaccccat cttctgcacc atcgtgctgg
     4081 gcacctttgc catccagata gtgatcgtgc agtttggagg gaagccattc agctgctctc
     4141 cactgcagct ggaccagtgg atgtggtgca tattcattgg gttaggagag ctcgtttggg
     4201 gccaggtcat cgccaccatc ccgaccagca gactcaagtt cctcaaggag gcaggcaggc
     4261 tcacacagaa ggaggagatc ccggaggagg agctcaacga ggacgtggag gagatcgacc
     4321 acgcggagcg ggagctgcgg cggggccaga tcctgtggtt ccgaggcctg aatcggatcc
     4381 agacacagat ccgcgtcgtg aaggcgttcc gtagctctct ctatgaaggt ttagaaaagc
     4441 ctgaatctcg aacctccatc cataacttca tggctcatcc tgaattccgg atcgaagatt
     4501 cccagcccca catccccctc attgatgaca ccgacctgga agaagatgcc gcgctcaagc
     4561 agaactcgag cccgccgtca tccctcaaca agaacaacag cgccatcgac agtgggatca
     4621 acctgacgac cgacacaagc aaatcagcta cctcttcaag tccagggagc cccatccaca
     4681 gcctggagac gtcgctttag gcggccgccg actagtgagc tcgtcgaccc gggaattccg
     4741 gaccggtacc gaattccggg gggggggggg ggggggacag ctcagggctg cgatttcgcg
     4801 ccaaacttga cggcaatcct agcgtgaagg ctggtaggat tttatccccg ctgccatcat
     4861 ggttcgacca ttgaactgca tcgtcgccgt gtcccaaaat atggggattg gcaagaacgg
     4921 agacctaccc tggcctccgc tcaggaacga gttcaagtac ttccaaagaa tgaccacaac
     4981 ctcttcagtg gaaggtaaac agaatctggt gattatgggt aggaaaacct ggttctccat
     5041 tcctgagaag aatcgacctt taaaggacag aattaatata gttctcagta gagaactcaa
     5101 agaaccacca cgaggagctc attttcttgc caaaagtttg gatgatgcct taagacttat
     5161 tgaacaaccg gaattggcaa gtaaagtaga catggtttgg atagtcggag gcagttctgt
     5221 ttaccaggaa gccatgaatc aaccaggcca cctcagactc tttgtgacaa ggatcatgca
     5281 ggaatttgaa agtgacacgt ttttcccaga aattgatttg gggaaatata aacttctccc
     5341 agaataccca ggcgtcctct ctgaggtcca ggaggaaaaa ggcatcaagt ataagtttga
     5401 agtctacgag aagaaagact aacaggaaga tgctttcaag ttctctgctc ccctcctaaa
     5461 gctatgcatt ttttataaga ccatgggact tttgctggct ttagatcata atcagccata
     5521 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5581 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5641 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5701 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccccggc caacggtctg
     5761 gtgacccggc tgcgagagct cggtgtacct gagacgcgag taagcccttg agtcaaagac
     5821 gtagtcgttg caagtccgca ccaggtactg atatcccacc aaaaagtgcg gcggcggctg
     5881 gcggtagagg ggccagcgta gggtggccgg ggctccgggg gcgaggtctt ccaacataag
     5941 gcgatgatat ccgtagatgt acctggacat ccaggtgatg ccggcggcgg tggtggaggc
     6001 gcgcggaaag tcgcggacgc ggttccagat gttgcgcagc ggcaaaaagt gctccatggt
     6061 cgggacgctc tggccggtga ggcgtgcgca gtcgttgacg ctctagaccg tgcaaaagga
     6121 gagcctgtaa gcgggcactc ttccgtggtc tggtggataa attcgcaagg gtatcatggc
     6181 ggacgaccgg ggttcgaacc ccggatccgg ccgtccgccg tgatccatcc ggttaccgcc
     6241 cgcgtgtcga acccaggtgt gcgacgtcag acaacggggg agcgctcctt ttggcttcct
     6301 tccaggcgcg gcggctgctg cgctagcttt tttggcgagc tcgaattaat tctgcattaa
     6361 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6421 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6481 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6541 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6601 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6661 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6721 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6781 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6841 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6901 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6961 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7021 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7081 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7141 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7201 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7261 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7321 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7381 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7441 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7501 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7561 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7621 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7681 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7741 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7801 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7861 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7921 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7981 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8041 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8101 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8161 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8221 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8281 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8341 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8401 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     8461 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     8521 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     8581 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     8641 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     8701 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     8761 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct t
//
