LOCUS       sequence-356971-        8357 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 8357)
  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 8357)
  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: EGFP-hPMCA4b
FEATURES             Location/Qualifiers
     source          1..8357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4969..5021
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    5145..5266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5182..5201)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5236..5255
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5273..5728)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5410..5431)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5622..5641
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5755..5859
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5861..6218
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5861..6052
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5884..5904)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6069..6204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6253..7047
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6253..7047
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6307..6326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6917..6936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7235..7254)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7279..7326
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7655..8243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8144..8163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agaacgaacc catcagaccg tgtcttgcct
     1381 gccaactcga tggccgagag ccgtgaaggg gactttggct gcacagtaat ggaactgagg
     1441 aagctcatgg agctgcgttc gagggatgca ctgacccaga ttaatgtcca ctatggaggt
     1501 gtacagaatc tctgcagtag actgaaaacc tcccctgtgg aaggtctgtc tgggaaccct
     1561 gcagatctgg agaaacgtag gcaggtgttt ggacacaacg tgatcccccc caaaaagccc
     1621 aagactttct tagaattagt gtgggaagct cttcaagatg tcacgctcat catcctggag
     1681 attgcagcca tcatctccct ggtcctgtcc ttttatcgcc ctgctggtga agaaaatgaa
     1741 ctgtgtggtc aagtcgcaac taccccagaa gatgaaaatg aggcacaagc tggctggatt
     1801 gagggggcag ccatcctttt ctcagtgatc atcgtggtgt tagtgactgc ctttaatgat
     1861 tggagcaaag agaagcaatt ccgggggctg cagtgccgca ttgaacagga gcaaaagttc
     1921 tccatcatcc gaaacggtca actcatccag ctccctgtgg ctgagattgt ggttggtgat
     1981 attgcccaag tcaaatacgg tgatctgctg cctgcagatg gaatcctgat ccaagggaat
     2041 gatctgaaga ttgatgagag ctctctgaca ggggaatctg accatgtcaa gaagtccctg
     2101 gacaaagacc ccatgttgct ctcagggacc catgtcatgg aaggttctgg ccggatggtg
     2161 gtgacagctg ttggtgtcaa ctctcagact ggaatcatcc ttactctctt gggggtcaat
     2221 gaggatgacg aaggggagaa aaagaagaaa ggtaaaaaac aaggagtccc tgaaaatcgc
     2281 aacaaagcaa agacccaaga cggagtggcc ctggaaatcc agccactcaa cagccaggag
     2341 ggaatcgaca atgaggaaaa ggacaagaag gcagtcaagg tgcctaaaaa ggagaagtca
     2401 gtgctgcagg gcaagctgac tcgcctggct gttcagattg ggaaagccgg tctgctcatg
     2461 tctgctctca cggttttcat cctgattcta tactttgtga ttgacaactt tgtgataaat
     2521 cgcagaccat ggctccctga gtgtactccc atctacatcc agtactttgt caagttcttc
     2581 atcatcggca tcactgtact ggtggtggct gtgccagagg ggctgcctct ggctgtcacc
     2641 atctcgctgg cctactctgt gaagaaaatg atgagagaca ataacctagt acggcacttg
     2701 gatgcttgtg agaccatggg caacgccacc gccatctgct ctgataagac aggcacgttg
     2761 accatgaacc gcatgactgt ggtacaagct tatattgggg gcatccatta ccgtcaaatc
     2821 ccaagccctg atgtcttcct gcccaaagtc ctggacctca ttgtcaatgg catttctatc
     2881 aacagtgctt atacctccaa gattctgcct ccagagaagg agggaggcct gcctcggcag
     2941 gtgggcaaca agaccgagtg tgctctgcta ggctttgtca cagatctgaa gcaggattat
     3001 caggctgtgc gtaatgaagt gcccgaggag aagctctaca aggtgtacac ctttaactca
     3061 gtgcgcaagt caatgagcac cgtcatcagg aatcccaacg gtggcttccg tatgtacagc
     3121 aagggcgcct ctgagatcat cttgcgcaag tgtaatcgaa tcctggaccg gaaaggggaa
     3181 gcagtgccat tcaagaataa agacagagat gatatggtac gcactgtcat cgagcccatg
     3241 gcctgtgatg gactccggac tatctgcata gcttaccggg acttcgatga cacagagccc
     3301 tcttgggaca atgagaatga gatcctcacc gaactgacct gtatcgcggt ggtgggcatt
     3361 gaggaccttg tgcgcccaga ggtgccagat gctattgcca aatgcaaaca agctggcatt
     3421 actgtcagaa tggtgacagg tgacaacatc aacacagccc gggccattgc caccaaatgt
     3481 ggcattctga cacctgggga tgacttcctg tgcttagaag gcaaagaatt caaccggctc
     3541 atccgcaacg agaaaggcga ggtagagcaa gaaaagctgg acaagatctg gcctaagctt
     3601 cgggtcctgg cgcgatcttc tcccactgac aagcacaccc tggtgaaagg cataattgac
     3661 agcactgttg gggaacaccg gcaggtcgtg gctgtcactg gtgatggcac aaatgacggg
     3721 cctgctctga agaaagcgga tgttggtttt gccatgggca tcgcaggcac agatgtagca
     3781 aaggaggctt cagacatcat cctaacagat gacaacttca ccagcattgt gaaggcagtg
     3841 atgtggggac gaaatgtcta tgacagcatc tccaagttcc tgcagttcca gctcactgtc
     3901 aatgtggtgg ccgtgattgt agccttcact ggagcctgta tcactcagga ttccccattg
     3961 aaagctgtgc agatgttgtg ggttaatctg atcatggaca cttttgcttc attggccctg
     4021 gccacagagc cccctacgga atctctgttg aagcggcgcc cctatggccg aaataagcct
     4081 ctgatctcac gcactatgat gaagaacatc ttgggccatg cattctatca gctcattgtc
     4141 atctttatcc ttgtctttgc gggtgagaaa ttctttgata ttgatagtgg gaggaaggca
     4201 cctctacatt caccacccag ccagcactat accattgttt ttaacacctt cgtgctgatg
     4261 cagctcttca atgaaatcaa ctcccgaaag atccatggag agaagaacgt cttttcaggc
     4321 atctaccgca acattatctt ctgctctgta gtcttgggca cattcatctg ccagattttc
     4381 atcgtggaat ttgggggtaa acccttcagt tgtacaagcc tcagcctgtc tcagtggctg
     4441 tggtgtctct tcattgggat tggagaactt ctgtggggcc agttcatctc cgcaatacct
     4501 acccgatccc tgaagttcct gaaggaggct gggcatggca ccaccaaaga ggagatcacc
     4561 aaggatgccg agggactgga tgagattgac catgctgaga tggagctgcg ccgaggccag
     4621 atcctctggt tccggggcct gaaccggatc cagactcaga tcaaagtggt caaagcgttc
     4681 catagttccc tccacgaaag cattcagaaa ccctacaacc aaaagtccat ccacagcttc
     4741 atgacccacc ctgaattcgc catagaggag gagttgccac gaacaccact cctggatgag
     4801 gaagaggagg aaaatcctga caaggcttct aagtttggga ctagggtgct cctgttggat
     4861 ggtgaggtca ctccatatgc caatacaaac aacaatgcgg tggattgcaa ccaagtgcag
     4921 ctcccccagt cggacagctc tctacagagc ctagagacat cagtttgact cgagctcaag
     4981 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     5041 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5101 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     5281 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5341 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5401 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5461 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5521 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5581 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5641 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5701 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5821 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5881 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5941 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6061 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6121 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6181 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     6241 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6301 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6361 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6421 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6481 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6541 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6601 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6661 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6721 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6781 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6841 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6901 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6961 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7021 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     7081 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     7141 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     7201 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     7261 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     7321 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     7381 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     7441 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     7501 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7561 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7621 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7681 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7741 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7801 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7861 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7921 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7981 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8041 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8101 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8161 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8221 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8281 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8341 gtattaccgc catgcat
//
