LOCUS       sequence-67920-e        8837 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 8837)
  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 8837)
  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-hPMCA2x/b
FEATURES             Location/Qualifiers
     source          1..8837
                     /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      1084..1093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1591..1602)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2059..2068
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4885..5448
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    5636..5770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5673..5692)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5727..5746
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6443..6460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6614..7202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6694..6713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7373..8164,8165..8233))
                     /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     7996..8015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8234..8338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8406..8424
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8462..8484)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8584..8603
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8797..8819
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8811..8828
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8812..8828
                     /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 gtcgacgagc tcactagtcg
     1081 gcggccgcca tgggtgacat gaccaacagc gacttttact ccaaaaacca aagaaatgag
     1141 tcgagccatg ggggcgagtt cgggtgcaca atggaggagc tccgctccct catggagctg
     1201 cggggcactg aggctgtggt caagatcaag gagacttatg gggacaccga agccatctgc
     1261 cggcgcctca aaacctcacc tgttgaaggt ttgccgggca ccgctccaga cctggaaaag
     1321 agaaagcaaa tttttgggca aaactttata cctccaaaga agccaaaaac cttcctgcag
     1381 ctcgtgtggg aggcgctgca ggacgtgacg ctcatcatcc tggagattgc cgccatcatc
     1441 tccctggggc tgtccttcta ccacccgccc ggcgagggca acgaaggatg tgcgacggcc
     1501 cagggtgggg cagaggatga aggagaggca gaggcaggtt ggatcgaggg ggccgccatt
     1561 ctcctctcag ttatctgtgt ggtcctggtc acggccttca atgactggag caaagagaaa
     1621 cagttccggg gcctgcagag ccgcatcgag caggaacaga aatttaccgt ggtccgggct
     1681 ggccaggtgg tgcagatccc tgtggctgag atcgtggttg gggacatagc ccaggtcaaa
     1741 tatggtgacc tcctccctgc cgacggcctc ttcatccagg gcaatgacct caagattgat
     1801 gaaagctccc taactggaga gtctgaccag gtgcgcaagt ccgtggacaa ggaccccatg
     1861 ctgctgtcag gaacccacgt gatggagggc tcaggacgga tgttggtgac tgctgtgggt
     1921 gtgaactctc agactggcat catctttacc ctcctggggg ctggtggtga agaggaagag
     1981 aagaaagaca aaaaaggtaa aatgcaggat ggcaatgtgg acgccagcca gagcaaagcc
     2041 aaacaacagg acggggcagc cgccatggag atgcagcccc tcaagagtgc cgagggcggc
     2101 gacgctgacg acaggaagaa ggccagcatg cacaagaagg agaagtccgt gctgcagggc
     2161 aagctcacca agctggctgt gcagatcggg aaggcgggct tggtgatgtc agccatcacg
     2221 gtgatcatcc tggtgctcta cttcactgtg gacaccttcg tggtcaacaa gaagccgtgg
     2281 ctgcctgagt gcacgcccgt ctacgtgcag tactttgtca agttcttcat cattggcgtg
     2341 acggtgctgg tggtcgccgt gcccgagggg ctccctctgg ccgtcaccat ctcgttggcc
     2401 tattcggtga agaaaatgat gaaggacaac aacctggtac gccacctgga tgcctgtgag
     2461 accatgggca atgccacagc catctgctca gacaagacag gcacgctgac caccaatcgc
     2521 atgacagtgg tacaggccta tgtcggcgac gtccactata aagagatccc cgaccccagc
     2581 tccatcaaca ccaagaccat ggagctgctg atcaatgcca tcgccatcaa cagcgcctac
     2641 accaccaaga ttctgccccc agagaaggag ggcgccctgc ctcggcaggt gggcaacaag
     2701 acggagtgcg gcctgctggg cttcgtgctg gacctgaagc aggactacga gcccgtgcgc
     2761 agccagatgc cagaggagaa gttgtacaaa gtgtacacct tcaactccgt gcgcaagtcc
     2821 atgagcactg tcatcaagct gcccgacgag agcttccgca tgtacagcaa gggggcttct
     2881 gagatcgtgc tcaagaagtg ctgcaaaatc ctcaatgggg cgggagagcc tcgtgtcttc
     2941 cggccccgcg accgggacga gatggtaaag aaggtgattg agcccatggc ttgcgatggg
     3001 ctccgcacta tctgcgtggc ctaccgcgac ttccccagca gcccggagcc ggactgggac
     3061 aatgagaatg acatcctcaa cgaactcacc tgcatctgcg tggtgggcat cgaggacccg
     3121 gtgcggccag aggtcccaga agccatccgc aagtgccagc gggcaggcat cacggtccgc
     3181 atggtcactg gcgacaatat caacacggct cgggccatcg ccatcaagtg tggcatcatc
     3241 catcctgggg aggactttct gtgcctcgag ggcaaggagt tcaacaggag gatccgcaac
     3301 gagaaggggg agattgagca ggagcgaatt gacaagatct ggccaaagct gcgggtgctg
     3361 gctcgctcct ccccaacgga caagcatacc ctggttaaag gcatcatcga cagcacacac
     3421 actgagcagc ggcaggtggt ggccgtgacg ggggacggga ccaacgacgg gcctgcactc
     3481 aagaaggccg acgtgggctt cgccatgggc atcgcaggca ctgacgtggc caaggaggcc
     3541 tcagacatca tcctgacaga cgacaatttc agcagcatcg tcaaggcagt gatgtggggc
     3601 cgcaacgtct atgacagcat ctccaaattc ttgcagttcc agctcaccgt caacgtggtg
     3661 gccgtgattg tggccttcac aggcgcctgc atcacgcagg actcccctct gaaggccgtg
     3721 cagatgctct gggtgaacct catcatggac acgtttgcct cgctggcact ggccactgag
     3781 ccgcccacgg agaccctgct gctgaggaag ccgtacggcc gcaacaagcc gctcatctcc
     3841 aggaccatga tgaagaacat cctgggccat gctgtctacc agcttgccct catcttcacc
     3901 ctgctctttg ttggcgagaa gatgttccag atcgacagcg ggaggaacgc gcccctgcat
     3961 tcgccaccct cagaacatta caccatcatc ttcaacacct tcgtcatgat gcagctcttc
     4021 aacgagatca acgcccgcaa gatccacggc gagcgcaatg tctttgacgg catcttccgg
     4081 aaccccatct tctgcaccat cgtgctgggc acctttgcca tccagatagt gatcgtgcag
     4141 tttggaggga agccattcag ctgctctcca ctgcagctgg accagtggat gtggtgcata
     4201 ttcattgggt taggagagct cgtttggggc caggtcatcg ccaccatccc gaccagcaga
     4261 ctcaagttcc tcaaggaggc aggcaggctc acacagaagg aggagatccc ggaggaggag
     4321 ctcaacgagg acgtggagga gatcgaccac gcggagcggg agctgcggcg gggccagatc
     4381 ctgtggttcc gaggcctgaa tcggatccag acacagatcc gcgtcgtgaa ggcgttccgt
     4441 agctctctct atgaaggttt agaaaagcct gaatctcgaa cctccatcca taacttcatg
     4501 gctcatcctg aattccggat cgaagattcc cagccccaca tccccctcat tgatgacacc
     4561 gacctggaag aagatgccgc gctcaagcag aactcgagcc cgccgtcatc cctcaacaag
     4621 aacaacagcg ccatcgacag tgggatcaac ctgacgaccg acacaagcaa atcagctacc
     4681 tcttcaagtc cagggagccc catccacagc ctggagacgt cgctttaggc ggccgctcta
     4741 gaggatccaa gcttacgtac gcgtatcgat ggtaccgaat tccggggggg gggggggggg
     4801 ggacagctca gggctgcgat ttcgcgccaa acttgacggc aatcctagcg tgaaggctgg
     4861 taggatttta tccccgctgc catcatggtt cgaccattga actgcatcgt cgccgtgtcc
     4921 caaaatatgg ggattggcaa gaacggagac ctaccctggc ctccgctcag gaacgagttc
     4981 aagtacttcc aaagaatgac cacaacctct tcagtggaag gtaaacagaa tctggtgatt
     5041 atgggtagga aaacctggtt ctccattcct gagaagaatc gacctttaaa ggacagaatt
     5101 aatatagttc tcagtagaga actcaaagaa ccaccacgag gagctcattt tcttgccaaa
     5161 agtttggatg atgccttaag acttattgaa caaccggaat tggcaagtaa agtagacatg
     5221 gtttggatag tcggaggcag ttctgtttac caggaagcca tgaatcaacc aggccacctc
     5281 agactctttg tgacaaggat catgcaggaa tttgaaagtg acacgttttt cccagaaatt
     5341 gatttgggga aatataaact tctcccagaa tacccaggcg tcctctctga ggtccaggag
     5401 gaaaaaggca tcaagtataa gtttgaagtc tacgagaaga aagactaaca ggaagatgct
     5461 ttcaagttct ctgctcccct cctaaagcta tgcatttttt ataagaccat gggacttttg
     5521 ctggctttag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5581 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5641 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5701 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5761 tgtctggatc cccggccaac ggtctggtga cccggctgcg agagctcggt gtacctgaga
     5821 cgcgagtaag cccttgagtc aaagacgtag tcgttgcaag tccgcaccag gtactgatat
     5881 cccaccaaaa agtgcggcgg cggctggcgg tagaggggcc agcgtagggt ggccggggct
     5941 ccgggggcga ggtcttccaa cataaggcga tgatatccgt agatgtacct ggacatccag
     6001 gtgatgccgg cggcggtggt ggaggcgcgc ggaaagtcgc ggacgcggtt ccagatgttg
     6061 cgcagcggca aaaagtgctc catggtcggg acgctctggc cggtgaggcg tgcgcagtcg
     6121 ttgacgctct agaccgtgca aaaggagagc ctgtaagcgg gcactcttcc gtggtctggt
     6181 ggataaattc gcaagggtat catggcggac gaccggggtt cgaaccccgg atccggccgt
     6241 ccgccgtgat ccatccggtt accgcccgcg tgtcgaaccc aggtgtgcga cgtcagacaa
     6301 cgggggagcg ctccttttgg cttccttcca ggcgcggcgg ctgctgcgct agcttttttg
     6361 gcgagctcga attaattctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6781 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7021 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8401 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8461 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8521 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8581 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8641 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8701 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8761 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     8821 cggccagtgc caagctt
//
