LOCUS       sequence-424686-        7099 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 7099)
  AUTHORS   Qin Y, Dittmer PJ, Park JG, Jansen KB, Palmer AE
  TITLE     Measuring steady-state and dynamic endoplasmic reticulum and Golgi 
            Zn2+ with genetically encoded sensors.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 May 3;108(18):7351-6. Epub 2011 Apr 
            18.
  PUBMED    21502528
REFERENCE   2  (bases 1 to 7099)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2011-05-3"; volume: "108(18)"; pages: 
            "7351-6. Epub 2011 Apr 18"
COMMENT     Sequence Label: pcDNA-ER-ZapCY1
FEATURES             Location/Qualifiers
     source          1..7099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        142..521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        522..725
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     676..696
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     770..789
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        770..788
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(873..875,876..878,879..1391)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             join(1779..1781,1782..1784,1785..2495)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGKDEL*K"
     promoter        complement(2559..2577)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2560..2577)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2597..2614)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2603..2827
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2873..3301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2960..2979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3170..3191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3310..3330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3315..3644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3495..3630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3557..3576
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3711..4505
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3765..3784)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4375..4394
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4679..4800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4716..4735)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4770..4789
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4849..4865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4849..4865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4862..4884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4873..4889
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4897..4903,4904..4921,4922..4927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4942..4963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5080..5097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5251..5839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5331..5350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6010..6801,6802..6870))
                     /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     6633..6652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6871..6975)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7050..7069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
       61 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      121 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      181 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      241 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      301 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      361 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      421 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      481 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      541 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      601 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      661 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      721 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      781 actataggga gacccaagct tgcggccgcc acatgctgct gcccgtcccc ctgctgctgg
      841 gcctgctggg cgccgccgcc gacggggatc ccatggtgag caagggcgag gagctgttca
      901 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aggttcagcg
      961 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1021 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc
     1081 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1141 ccgaaggcta cgtccaggag cgtaccatct tcttcaagga cgacggcaac tacaagaccc
     1201 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1261 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca
     1321 acgtctatat caccgccgac aagcagaaga acggcatcaa ggcccacttc aagatccgcc
     1381 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1441 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1501 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgcccgca
     1561 tgcataaaaa caatgactta aaatgcaaat ggaaagaatg tcctgagtct tgtagctcac
     1621 tatttgacct acaaagacat cttttgaagg atcatgtctc tcaagatttc aaacacccta
     1681 tggaaccatt agcttgtaat tgggaggact gtgatttcct tggagatgat acatgttcca
     1741 tagtgaacca tattaattgt caacatggta tcgagctcat ggtgagcaag ggcgaggagc
     1801 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1861 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1921 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     1981 gcctgatgtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2041 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2101 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2161 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2221 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2281 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2341 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc
     2401 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2461 ccgggatcac tctcggcaag gacgagctct aaaagtaaga attctgcaga tatccatcac
     2521 actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     2581 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2641 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2701 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2761 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2821 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2881 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2941 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3001 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3061 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3121 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3181 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3241 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3301 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3361 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3421 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3481 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3541 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3601 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3661 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3721 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3781 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3841 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3901 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3961 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4021 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4081 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4141 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4201 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4261 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4321 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4381 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4441 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4501 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4561 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4621 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4681 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4741 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4801 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5101 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5161 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5221 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5281 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5341 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5701 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5761 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5821 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5881 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5941 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6001 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6061 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6121 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6181 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6241 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6301 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6361 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6421 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6481 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6541 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6601 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6661 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6721 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6781 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6841 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6901 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6961 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7021 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7081 cagtatctgc tccctgctt
//
