LOCUS       sequence-345094-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cell surface localized zinc sensor with ~ 300 nM Kd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Chen M, Zhang S, Xing Y, Li X, He Y, Wang Y, Oberholzer J, Ai HW
  TITLE     Genetically Encoded, Photostable Indicators to Image Dynamic Zn(2+) 
            Secretion of Pancreatic Islets.
  JOURNAL   Anal Chem. 2019 Oct 1;91(19):12212-12219. doi: 
            10.1021/acs.analchem.9b01802. Epub 2019 Sep 10.
  PUBMED    31475537
REFERENCE   2  (bases 1 to 6084)
  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.9b01802"; journalName: "Anal Chem"; date: 
            "2019-10-1- 1"; volume: "91"; issue: "19"; pages: "12212-12219"
COMMENT     Sequence Label: pDisplay-ZIBG2
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..251
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(83..103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     330..349
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             448..1242
                     /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
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(502..521)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1112..1131
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1430..1449)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1474..1521
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2339..2358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2570..2794
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2783..2800
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2822..2968)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(2972..3001)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     3165..3184
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3424..3445
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3551..3572)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3839..3898)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     complement(3978..3997)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3979..3997)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4042..4245)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4071..4091)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4246..4625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4637..5092)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4774..4795)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4986..5005
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5119..5223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5224..5292,5293..6084)
                     /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(5442..5461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctgtcagacc aagtttactc atatatactt tagattgatt tattgaaaaa ggaagagtcc
       61 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
      121 tccccagcag gcagaagtat gcaaagcaag catctcaatt agtcagcaac caggtgtgga
      181 aagtccccag gctccccagc aggcagaagt atgcaaagca agcatctcaa ttagtcagca
      241 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
      301 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc ccggcctctg
      361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
      421 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
      481 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      541 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      601 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
      661 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      721 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      781 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      841 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      901 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
      961 gccaggctca aggcgagcaa gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     1021 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1081 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1141 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     1201 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     1261 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     1321 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     1381 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     1441 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1501 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1561 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1621 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1681 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1741 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1801 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1861 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1921 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1981 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2041 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2101 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2161 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2221 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2281 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2341 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2401 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2461 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2521 attctgtgga taaccgtatt accgccactg gttctttccg cctcagaagc catagagccc
     2581 accgcatccc cagcacgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     2641 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     2701 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     2761 caaacaacag atggctggca actagaaggc acagtcgagg ctgatctcga gcggccgcct
     2821 aacgtggctt cttctgccaa agcatgatga ggatgataag ggagatgatg gtgagcacca
     2881 ccagggccag gatggctgag atcaccacca ccttaaaggg caaggagtgt ggcaccacga
     2941 tgacctcctg cgtgtcctgg cccacagcat tcagatcctc ttctgagatg agtttttgtt
     3001 cgtcgacgtc agtcagctct gctccacaca cagggcactt gcctttagca gacgggttgt
     3061 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3121 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3181 cgtccttgaa gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga
     3241 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     3301 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3361 tcagcttgcc gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt
     3421 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     3481 tgctcaccat gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc
     3541 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg
     3601 aaagtttgga ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     3661 tgggggtgtt ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc
     3721 ggatcttgaa gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacacaat
     3781 ctgttaattc agcaccgcat actggacatt ttccctttgc gtcaaccata gatctgtcac
     3841 cagtggaacc tggaacccag agcagcagta cccatagcag gagtgtgtct gtctccatgg
     3901 tggatatccc caagccgaat tccagcacac tggcggccgt tactagtgga tccgagttcg
     3961 gtaccaagct tgggtctccc tatagtgagt cgtattaatt tcgataagcc agtaagcagt
     4021 gggttctcta gttagccaga gagttctgct tatatagacc tcccaccgta cacgcctacc
     4081 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     4141 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     4201 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
     4261 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     4321 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     4381 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     4441 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     4501 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     4561 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tcaataatca
     4621 atgtcaacgc gcgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4681 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4741 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4801 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4861 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4921 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4981 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5041 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     5101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5461 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6061 gcctcactga ttaagcattg gtaa
//
