LOCUS       sequence-424763-        7238 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 7238)
  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 7238)
  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-Golgi-ZapCY1
FEATURES             Location/Qualifiers
     source          1..7238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..493
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        494..697
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     648..668
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     742..761
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        742..760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1012..1014,1015..1017,1018..1530)
                     /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(1918..1920,1921..1923,1924..2634)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2708..2725)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2714..2938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2984..3412
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3071..3090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3281..3302
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3421..3441)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3426..3755
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3606..3741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3668..3687
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3822..4616
                     /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(3876..3895)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4486..4505
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4790..4911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4827..4846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4881..4900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4960..4976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4960..4976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4973..4995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4984..5000
                     /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(5008..5014,5015..5032,5033..5038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5053..5074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5191..5208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5362..5950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5442..5461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6121..6912,6913..6981))
                     /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     6744..6763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6982..7086)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7161..7180)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
       61 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      121 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      181 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      241 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      301 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      361 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      421 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      481 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      541 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      601 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      661 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      721 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg
      781 tttaaactta agcttgccac catgaggctt cgggagccgc tcctgagcgg cagcgccgcg
      841 atgccaggcg cgtccctaca gcgggcctgc cgcctgctcg tggccgtctg cgctctgcac
      901 cttggcgtca ccctcgttta ctacctggct ggccgcgacc tgagccgcct gccccaactg
      961 gtcggagtct ccacaccgct gcagtcgacg gtaccgcggg cccgggatcc catggtgagc
     1021 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1081 aacggccaca ggttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1141 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1201 accctgacct ggggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1261 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gtaccatctt cttcaaggac
     1321 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1381 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1441 tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1501 gcccacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1561 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1621 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1681 ttcgtgaccg ccgcccgcat gcataaaaac aatgacttaa aatgcaaatg gaaagaatgt
     1741 cctgagtctt gtagctcact atttgaccta caaagacatc ttttgaagga tcatgtctct
     1801 caagatttca aacaccctat ggaaccatta gcttgtaatt gggaggactg tgatttcctt
     1861 ggagatgata catgttccat agtgaaccat attaattgtc aacatggtat cgagctcatg
     1921 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1981 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2041 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2101 gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag
     2161 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2221 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2281 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2341 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2401 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2461 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2521 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2581 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctata caagtaagaa
     2641 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     2701 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2761 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2821 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2881 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2941 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3001 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3061 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3121 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3181 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3241 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3301 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3361 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3421 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3481 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3601 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3661 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3721 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3781 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3841 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3901 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3961 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4021 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4081 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4141 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4201 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4261 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4321 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4381 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4441 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4501 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4561 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4621 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4681 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4741 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4801 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4861 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4921 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4981 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5041 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5101 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5161 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5221 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5281 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5341 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5401 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5461 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5521 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5581 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5641 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5701 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5761 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5821 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5881 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6181 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6241 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6301 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6361 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6421 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6481 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6541 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6601 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6661 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6721 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6781 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6841 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6901 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6961 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7021 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7081 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7141 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7201 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgc
//
