LOCUS       sequence-67923-e        8478 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 8478)
  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 8478)
  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-hPCM2w/b delta6
FEATURES             Location/Qualifiers
     source          1..8478
                     /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"
     regulatory      1390..1399
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1897..1908)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2458..2467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5266..5387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5303..5322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5357..5376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5394..5849)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5531..5552)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5743..5762
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5876..5980
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5982..6339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5982..6173
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6005..6025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6190..6325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6374..7168
                     /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             6374..7168
                     /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(6428..6447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7038..7057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7356..7375)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7400..7447
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7776..8364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8265..8284
                     /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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcggtggcgg ccgccatggg tgacatgacc aacagcgact tttactccaa aaaccaaaga
     1441 aatgagtcga gccatggggg cgagttcggg tgcacaatgg aggagctccg ctccctcatg
     1501 gagctgcggg gcactgaggc tgtggtcaag atcaaggaga cttatgggga caccgaagcc
     1561 atctgccggc gcctcaaaac ctcacctgtt gaaggtttgc cgggcaccgc tccagacctg
     1621 gaaaagagaa agcaaatttt tgggcaaaac tttatacctc caaagaagcc aaaaaccttc
     1681 ctgcagctcg tgtgggaggc gctgcaggac gtgacgctca tcatcctgga gattgccgcc
     1741 atcatctccc tggggctgtc cttctaccac ccgcccggcg agggcaacga aggatgtgcg
     1801 acggcccagg gtggggcaga ggatgaagga gaggcagagg caggttggat cgagggggcc
     1861 gccattctcc tctcagttat ctgtgtggtc ctggtcacgg ccttcaatga ctggagcaaa
     1921 gagaaacagt tccggggcct gcagagccgc atcgagcagg aacagaaatt taccgtggtc
     1981 cgggctggcc aggtggtgca gatccctgtg gctgagatcg tggttgggga catagcccag
     2041 gtcaaatatg gtgacctcct ccctgccgac ggcctcttca tccagggcaa tgacctcaag
     2101 attgatgaaa gctccctaac tggagagtct gaccaggtgc gcaagtccgt ggacaaggac
     2161 cccatgctgc tgtcaggaac ccacgtgatg gagggctcag gacggatgtt ggtgactgct
     2221 gtgggtgtga actctcagac tggcatcatc tttaccctcc tgggggctgg tggtgaagag
     2281 gaagagaaga aagacaaaaa aggtgtgaag aagggggatg gccttcagct accagcagca
     2341 gacggtgcgg cagcttcaaa cgctgcagat agtgcgaatg ccagcctagt caatggtaaa
     2401 atgcaggatg gcaatgtgga cgccagccag agcaaagcca aacaacagga cggggcagcc
     2461 gccatggaga tgcagcccct caagagtgcc gagggcggcg acgctgacga caggaagaag
     2521 gccagcatgc acaagaagga gaagtccgtg ctgcagggca agctcaccaa gctggctgtg
     2581 cagatcggga aggcgggctt ggtgatgtca gccatcacgg tgatcatcct ggtgctctac
     2641 ttcactgtgg acaccttcgt ggtcaacaag aagccgtggc tgcctgagtg cacgcccgtc
     2701 tacgtgcagt actttgtcaa gttcttcatc attggcgtga cggtgctggt ggtcgccgtg
     2761 cccgaggggc tccctctggc cgtcaccatc tcgttggcct attcggtgaa gaaaatgatg
     2821 aaggacaaca acctggtacg ccacctggat gcctgtgaga ccatgggcaa tgccacagcc
     2881 atctgctcag acaagacagg cacgctgacc accaatcgca tgacagtggt acaggcctat
     2941 gtcggcgacg tccactataa agagatcccc gaccccagct ccatcaacac caagaccatg
     3001 gagctgctga tcaatgccat cgccatcaac agcgcctaca ccaccaagat tctgccccca
     3061 gagaaggagg gcgccctgcc tcggcaggtg ggcaacaaga cggagtgcgg cctgctgggc
     3121 ttcgtgctgg acctgaagca ggactacgag cccgtgcgca gccagatgcc agaggagaag
     3181 ttgtacaaag tgtacacctt caactccgtg cgcaagtcca tgagcactgt catcaagctg
     3241 cccgacgaga gcttccgcat gtacagcaag ggggcttctg agatcgtgct caagaagtgc
     3301 tgcaaaatcc tcaatggggc gggagagcct cgtgtcttcc ggccccgcga ccgggacgag
     3361 atggtaaaga aggtgattga gcccatggct tgcgatgggc tccgcactat ctgcgtggcc
     3421 taccgcgact tccccagcag cccggagccg gactgggaca atgagaatga catcctcaac
     3481 gaactcacct gcatctgcgt ggtgggcatc gaggacccgg tgcggccaga ggtcccagaa
     3541 gccatccgca agtgccagcg ggcaggcatc acggtccgca tggtcactgg cgacaatatc
     3601 aacacggctc gggccatcgc catcaagtgt ggcatcatcc atcctgggga ggactttctg
     3661 tgcctcgagg gcaaggagtt caacaggagg atccgcaacg agaaggggga gattgagcag
     3721 gagcgaattg acaagatctg gccaaagctg cgggtgctgg ctcgctcctc cccaacggac
     3781 aagcataccc tggttaaagg catcatcgac agcacacaca ctgagcagcg gcaggtggtg
     3841 gccgtgacgg gggacgggac caacgacggg cctgcactca agaaggccga cgtgggcttc
     3901 gccatgggca tcgcaggcac tgacgtggcc aaggaggcct cagacatcat cctgacagac
     3961 gacaatttca gcagcatcgt caaggcagtg atgtggggcc gcaacgtcta tgacagcatc
     4021 tccaaattct tgcagttcca gctcaccgtc aacgtggtgg ccgtgattgt ggccttcaca
     4081 ggcgcctgca tcacgcagga ctcccctctg aaggccgtgc agatgctctg ggtgaacctc
     4141 atcatggaca cgtttgcctc gctggcactg gccactgagc cgcccacgga gaccctgctg
     4201 ctgaggaagc cgtacggccg caacaagccg ctcatctcca ggaccatgat gaagaacatc
     4261 ctgggccatg ctgtctacca gcttgccctc atcttcaccc tgctctttgt tggcgagaag
     4321 atgttccaga tcgacagcgg gaggaacgcg cccctgcatt cgccaccctc agaacattac
     4381 accatcatct tcaacacctt cgtcatgatg cagctcttca acgagatcaa cgcccgcaag
     4441 atccacggcg agcgcaatgt ctttgacggc atcttccgga accccatctt ctgcaccatc
     4501 gtgctgggca cctttgccat ccagatagtg atcgtgcagt ttggagggaa gccattcagc
     4561 tgctctccac tgcagctgga ccagtggatg tggtgcatat tcattgggtt aggagagctc
     4621 gtttggggcc aggtcatcgc caccatcccg accagcagac tcaagttcct caaggaggca
     4681 ggcaggctca cacagaagga ggagatcccg gaggaggagc tcaacgagga cgtggaggag
     4741 atcgaccacg cggagcggga gctgcggcgg ggccagatcc tgtggttccg aggcctgaat
     4801 cggatccaga cacagatccg cgtcgtgaag gcgttccgta gctctctcta tgaaggttta
     4861 gaaaagcctg aatctcgaac ctccatccat aacttcatgg ctcatcctga attccggatc
     4921 gaagattccc agccccacat ccccctcatt gatgacaccg acctggaaga agatgccgcg
     4981 ctcaagcaga actcgagccc gccgtcatcc ctcaacaaga acaacagcgc catcgacagt
     5041 gggatcaacc tgacgaccga cacaagcaaa tcagctacct cttcaagtcc agggagcccc
     5101 atccactagc ctggagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg
     5161 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     5221 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     5281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     5401 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5461 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5521 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5581 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5641 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5701 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5761 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5821 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     5881 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5941 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     6001 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     6061 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     6121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     6181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     6361 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6421 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6481 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6541 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6601 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6661 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6721 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6781 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6841 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6901 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6961 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     7021 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     7081 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7141 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     7201 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     7261 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7321 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     7381 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     7441 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     7501 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     7561 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     7621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7681 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7921 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8401 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8461 cgtattaccg ccatgcat
//
