LOCUS       sequence-345092-        6882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cytosolic ZnGreen1 zinc sensor with 633 nM Kd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6882)
  AUTHORS   Chen Z, Ai HW
  TITLE     Single Fluorescent Protein-Based Indicators for Zinc Ion (Zn2+).
  JOURNAL   Anal Chem. 2016 Sep 20;88(18):9029-36. doi: 
            10.1021/acs.analchem.6b01653. Epub 2016 Aug 29.
  PUBMED    27539450
REFERENCE   2  (bases 1 to 6882)
  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"; doi: 
            "10.1021/acs.analchem.6b01653"; journalName: "Anal Chem"; date: 
            "2016-09-20- 20"; volume: "88"; issue: "18"; pages: "9029-36"
COMMENT     Sequence Label: pcDNA3-ZnGreen1
FEATURES             Location/Qualifiers
     source          1..6882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    80..304
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     293..310
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        330..348
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     330..347
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    364..952
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             470..481
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     879..899
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     regulatory      1876..1885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1901..1920)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1902..1920)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1965..2168)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1994..2014)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2169..2548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2720..2739
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2814..2918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2919..2987,2988..3779)
                     /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     complement(3137..3156)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3950..4538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4439..4458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4692..4709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4826..4847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4862..4867,4868..4885,4886..4892)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4900..4916
                     /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)."
     primer_bind     4905..4927
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4924..4940
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4924..4940
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4989..5110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5000..5019)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5054..5073
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5284..6078)
                     /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(5395..5414)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6005..6024
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6145..6474)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6159..6294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6213..6232)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6459..6479
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(join(6488..6882,1..34))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6598..6619)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6810..6829
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg
       61 gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
      121 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
      181 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
      241 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
      301 acagtcgagg ctgatcagcg agctctagca tttaggtgac actatagaat agggccctct
      361 agagcgggga ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg
      421 gaagaggccg cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg
      481 aagggacgta gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc
      541 aaggaaagga cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag
      601 cccctgtcca gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg
      661 ggaaagcgaa agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg
      721 ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct
      781 cataaagaga cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag
      841 caatagcatg atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat
      901 agttaagaat accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttgtcgacct
      961 cgagttactt gtacagctcg tccatgccgt cggtggagtt gcgggccgcg gcgctctcgt
     1021 aaacggtcac cttgttgtag tccttgtcgt ggttcaggat ctcgatgtgg tggtccacaa
     1081 agtgatagtc gggcagcttc accgccttct tggccctgta gatggtcttg aagtcaacgc
     1141 ggtggtggcc gccgcccttc agcagcagct tgtgcttgac gtcgcccttc agcacgccgt
     1201 cgcgcacgta catcttctca tcacaagggt gctgacaatt gatgcggttg actatagagc
     1261 aggtgtcatc acccgggaaa tcacagtcct cccagttgca ggccagcggt tccatcggac
     1321 gtttagagtc ctgtgaaaca tgatctttga gaaggtgcct ctgcagatca aacaaggagg
     1381 agcagctctc tggacactcc ttccacttgc atttcaggtc attccccaac cagccggtgg
     1441 tcttcttctg catcacggga ccgttggggg ggaagttctc gcccttgagg tgtatctcgt
     1501 agatgaagga gccctcctcc atcgagatgt cggacttcac cttcacgatg cccttgtcct
     1561 cgaaggtcat ggtgcgctcc caagagtagc cctcggggaa ggactgcttg aagtagtcgg
     1621 ggatgtcgtc ggggtacttg gtgaaggccc tgttgccgta ggcgaacgcg gtggtcagaa
     1681 tgtcgtagga gaagggcagg ggggctccct ccttcacctc caggttgatg gtgtgggtgc
     1741 cgtcgtaggg cttgccctcg ccctcgccct cgatcacgaa ggcgtggcca ttcacgttgc
     1801 cctccatctt cagcttgatc ttcatgtcgg gcttgattac gcccattgtg gtctcctcgc
     1861 ccttgctcac gtcgaccatg gtggcggcaa gcttgggtct ccctatagtg agtcgtatta
     1921 atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct gcttatatag
     1981 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     2041 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     2101 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     2161 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     2221 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     2281 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     2341 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     2401 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     2461 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     2521 tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc
     2581 agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt
     2641 gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc
     2701 agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga
     2761 tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2821 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2881 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2941 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3001 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3061 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3121 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3181 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3241 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3301 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3361 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3421 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3481 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3541 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3601 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3661 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3721 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3781 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3841 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3901 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3961 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4021 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4081 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4141 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4201 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4261 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4321 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4381 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4501 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4561 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4621 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4681 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4741 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4801 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4861 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4921 acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     4981 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5041 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5101 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     5161 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     5221 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     5281 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     5341 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     5401 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     5461 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     5521 cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg
     5581 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     5641 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     5701 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     5761 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     5821 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     5881 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     5941 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     6001 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     6061 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atccgaaaat
     6121 ggatatacaa gctcccggga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     6181 ctacttctgg aatagctcag aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat
     6241 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     6301 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6361 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     6421 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     6481 cagaattaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     6541 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     6601 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     6661 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     6721 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     6781 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6841 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca ca
//
