LOCUS       sequence-67921-e        8930 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 8930)
  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 8930)
  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-hPMCA2w/b
FEATURES             Location/Qualifiers
     source          1..8930
                     /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      2152..2161
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4978..5541
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    5729..5863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5766..5785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5820..5839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6536..6553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6707..7295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6787..6806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7466..8257,8258..8326))
                     /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     8089..8108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8327..8431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8499..8517
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8555..8577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8677..8696
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8890..8912
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8904..8921
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8905..8921
                     /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 aaaaaggtgt gaagaagggg gatggccttc agctaccagc agcagacggt
     2041 gcggcagctt caaacgctgc agatagtgcg aatgccagcc tagtcaatgg taaaatgcag
     2101 gatggcaatg tggacgccag ccagagcaaa gccaaacaac aggacggggc agccgccatg
     2161 gagatgcagc ccctcaagag tgccgagggc ggcgacgctg acgacaggaa gaaggccagc
     2221 atgcacaaga aggagaagtc cgtgctgcag ggcaagctca ccaagctggc tgtgcagatc
     2281 gggaaggcgg gcttggtgat gtcagccatc acggtgatca tcctggtgct ctacttcact
     2341 gtggacacct tcgtggtcaa caagaagccg tggctgcctg agtgcacgcc cgtctacgtg
     2401 cagtactttg tcaagttctt catcattggc gtgacggtgc tggtggtcgc cgtgcccgag
     2461 gggctccctc tggccgtcac catctcgttg gcctattcgg tgaagaaaat gatgaaggac
     2521 aacaacctgg tacgccacct ggatgcctgt gagaccatgg gcaatgccac agccatctgc
     2581 tcagacaaga caggcacgct gaccaccaat cgcatgacag tggtacaggc ctatgtcggc
     2641 gacgtccact ataaagagat ccccgacccc agctccatca acaccaagac catggagctg
     2701 ctgatcaatg ccatcgccat caacagcgcc tacaccacca agattctgcc cccagagaag
     2761 gagggcgccc tgcctcggca ggtgggcaac aagacggagt gcggcctgct gggcttcgtg
     2821 ctggacctga agcaggacta cgagcccgtg cgcagccaga tgccagagga gaagttgtac
     2881 aaagtgtaca ccttcaactc cgtgcgcaag tccatgagca ctgtcatcaa gctgcccgac
     2941 gagagcttcc gcatgtacag caagggggct tctgagatcg tgctcaagaa gtgctgcaaa
     3001 atcctcaatg gggcgggaga gcctcgtgtc ttccggcccc gcgaccggga cgagatggta
     3061 aagaaggtga ttgagcccat ggcttgcgat gggctccgca ctatctgcgt ggcctaccgc
     3121 gacttcccca gcagcccgga gccggactgg gacaatgaga atgacatcct caacgaactc
     3181 acctgcatct gcgtggtggg catcgaggac ccggtgcggc cagaggtccc agaagccatc
     3241 cgcaagtgcc agcgggcagg catcacggtc cgcatggtca ctggcgacaa tatcaacacg
     3301 gctcgggcca tcgccatcaa gtgtggcatc atccatcctg gggaggactt tctgtgcctc
     3361 gagggcaagg agttcaacag gaggatccgc aacgagaagg gggagattga gcaggagcga
     3421 attgacaaga tctggccaaa gctgcgggtg ctggctcgct cctccccaac ggacaagcat
     3481 accctggtta aaggcatcat cgacagcaca cacactgagc agcggcaggt ggtggccgtg
     3541 acgggggacg ggaccaacga cgggcctgca ctcaagaagg ccgacgtggg cttcgccatg
     3601 ggcatcgcag gcactgacgt ggccaaggag gcctcagaca tcatcctgac agacgacaat
     3661 ttcagcagca tcgtcaaggc agtgatgtgg ggccgcaacg tctatgacag catctccaaa
     3721 ttcttgcagt tccagctcac cgtcaacgtg gtggccgtga ttgtggcctt cacaggcgcc
     3781 tgcatcacgc aggactcccc tctgaaggcc gtgcagatgc tctgggtgaa cctcatcatg
     3841 gacacgtttg cctcgctggc actggccact gagccgccca cggagaccct gctgctgagg
     3901 aagccgtacg gccgcaacaa gccgctcatc tccaggacca tgatgaagaa catcctgggc
     3961 catgctgtct accagcttgc cctcatcttc accctgctct ttgttggcga gaagatgttc
     4021 cagatcgaca gcgggaggaa cgcgcccctg cattcgccac cctcagaaca ttacaccatc
     4081 atcttcaaca ccttcgtcat gatgcagctc ttcaacgaga tcaacgcccg caagatccac
     4141 ggcgagcgca atgtctttga cggcatcttc cggaacccca tcttctgcac catcgtgctg
     4201 ggcacctttg ccatccagat agtgatcgtg cagtttggag ggaagccatt cagctgctct
     4261 ccactgcagc tggaccagtg gatgtggtgc atattcattg ggttaggaga gctcgtttgg
     4321 ggccaggtca tcgccaccat cccgaccagc agactcaagt tcctcaagga ggcaggcagg
     4381 ctcacacaga aggaggagat cccggaggag gagctcaacg aggacgtgga ggagatcgac
     4441 cacgcggagc gggagctgcg gcggggccag atcctgtggt tccgaggcct gaatcggatc
     4501 cagacacaga tccgcgtcgt gaaggcgttc cgtagctctc tctatgaagg tttagaaaag
     4561 cctgaatctc gaacctccat ccataacttc atggctcatc ctgaattccg gatcgaagat
     4621 tcccagcccc acatccccct cattgatgac accgacctgg aagaagatgc cgcgctcaag
     4681 cagaactcga gcccgccgtc atccctcaac aagaacaaca gcgccatcga cagtgggatc
     4741 aacctgacga ccgacacaag caaatcagct acctcttcaa gtccagggag ccccatccac
     4801 agcctggaga cgtcgcttta ggcggccgct ctagaggatc caagcttacg tacgcgtatc
     4861 gatggtaccg aattccgggg gggggggggg gggggacagc tcagggctgc gatttcgcgc
     4921 caaacttgac ggcaatccta gcgtgaaggc tggtaggatt ttatccccgc tgccatcatg
     4981 gttcgaccat tgaactgcat cgtcgccgtg tcccaaaata tggggattgg caagaacgga
     5041 gacctaccct ggcctccgct caggaacgag ttcaagtact tccaaagaat gaccacaacc
     5101 tcttcagtgg aaggtaaaca gaatctggtg attatgggta ggaaaacctg gttctccatt
     5161 cctgagaaga atcgaccttt aaaggacaga attaatatag ttctcagtag agaactcaaa
     5221 gaaccaccac gaggagctca ttttcttgcc aaaagtttgg atgatgcctt aagacttatt
     5281 gaacaaccgg aattggcaag taaagtagac atggtttgga tagtcggagg cagttctgtt
     5341 taccaggaag ccatgaatca accaggccac ctcagactct ttgtgacaag gatcatgcag
     5401 gaatttgaaa gtgacacgtt tttcccagaa attgatttgg ggaaatataa acttctccca
     5461 gaatacccag gcgtcctctc tgaggtccag gaggaaaaag gcatcaagta taagtttgaa
     5521 gtctacgaga agaaagacta acaggaagat gctttcaagt tctctgctcc cctcctaaag
     5581 ctatgcattt tttataagac catgggactt ttgctggctt tagatcataa tcagccatac
     5641 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     5701 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     5761 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5821 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccccggcc aacggtctgg
     5881 tgacccggct gcgagagctc ggtgtacctg agacgcgagt aagcccttga gtcaaagacg
     5941 tagtcgttgc aagtccgcac caggtactga tatcccacca aaaagtgcgg cggcggctgg
     6001 cggtagaggg gccagcgtag ggtggccggg gctccggggg cgaggtcttc caacataagg
     6061 cgatgatatc cgtagatgta cctggacatc caggtgatgc cggcggcggt ggtggaggcg
     6121 cgcggaaagt cgcggacgcg gttccagatg ttgcgcagcg gcaaaaagtg ctccatggtc
     6181 gggacgctct ggccggtgag gcgtgcgcag tcgttgacgc tctagaccgt gcaaaaggag
     6241 agcctgtaag cgggcactct tccgtggtct ggtggataaa ttcgcaaggg tatcatggcg
     6301 gacgaccggg gttcgaaccc cggatccggc cgtccgccgt gatccatccg gttaccgccc
     6361 gcgtgtcgaa cccaggtgtg cgacgtcaga caacggggga gcgctccttt tggcttcctt
     6421 ccaggcgcgg cggctgctgc gctagctttt ttggcgagct cgaattaatt ctgcattaat
     6481 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6541 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6601 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6661 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6721 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6781 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6841 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6901 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6961 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7021 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7081 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7141 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7201 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7261 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7321 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7381 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7441 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7501 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7561 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7621 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7681 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7741 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7801 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7861 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7921 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7981 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8041 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8101 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8161 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8221 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8281 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8341 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8401 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8461 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     8521 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     8581 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     8641 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     8701 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     8761 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     8821 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     8881 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
//
