LOCUS       sequence-352279-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Far-red fluorescent zinc indicator FRISZ in pTorPE vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  AUTHORS   Wu T, Kumar M, Zhang J, Zhao S, Drobizhev M, McCollum M, Anderson 
            CT, Wang Y, Pokorny A, Tian X, Zhang Y, Tzounopoulos T, Ai HW
  TITLE     A genetically encoded far-red fluorescent indicator for imaging 
            synaptically released Zn(2).
  JOURNAL   Sci Adv. 2023 Mar;9(9):eadd2058. doi: 10.1126/sciadv.add2058. Epub 
            2023 Mar 1.
  PUBMED    36857451
REFERENCE   2  (bases 1 to 5126)
  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: "Sci Adv."; 
            date: "2023-03"; volume: "9(9):eadd2058. doi"; pages: " 
            10.1126/sciadv.add2058. Epub 2023 Mar 1"
COMMENT     Sequence Label: pTorPE-FRISZ
FEATURES             Location/Qualifiers
     source          1..5126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             327..338
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             490..522
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(526..534,535..549)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1402..1425,1426..1461,1462..1485)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(1527..1544)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1697..1783
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1875..1902
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1921..2012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2013..2081,2082..2873)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2231..2250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3044..3632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3533..3552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3786..3803
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3818..3958
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3884..3903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4044..4066
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4222..5100)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gggtttaaag aggagaaagg tcatgaacaa taacgatctc
      361 tttcaggcat cacgtcggcg ttttctggca caactcggca gcttaaccga cgccgggacg
      421 ctggggccgt cattgttaac gccgcgacgt gcgactgcgg cgcaagcggc gactgacgct
      481 tctagaggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      541 gatgataagg atctcgccac actcgagatg gtcgacgcaa agggaaaatg tccagtatgc
      601 ggtgctgaat taacagatgt gtccaagaaa cccgcaaaga acctcaagat gccaggcgtc
      661 tattttattg acaggattct ggaaaggatc aaggaggctg gcaatgagac ctacgtcgag
      721 cagtacgagg tggctgtggc cagatactgc gaccacccta gcaaactggg gcacaaactt
      781 aaaggaggaa caggaggaag cagcgaggag ctgatcaagg agaacatgcg caccaagctg
      841 tacctgacgg gcaccgtgaa caaccactac ttcgaatgca ccgccgaagg ggagggccag
      901 ccctatgagg gcacccagac caacaggatt aaggtggtgc gcggaggccc cctgccgttc
      961 gcattcgaca tcctggcccc ctgctttatg tacgggagca agaccttcat taaccatcca
     1021 cctgacattc ccgattattt taagcagtcc ttccctgagg gcttcacatg ggagagaacc
     1081 accgttttcg aagacggggg cacactcact gctatacagg acaccagcct ccaggacggc
     1141 tgcctgatct acaacgtcca agtcagaggg gagaacttcc catccaacgg ccctgtgatg
     1201 cagaagaaaa cactcggctg ggaggccagt accgagaccc tgtaccccgc tgacggcagc
     1261 ctggaaggca gactgtactg ggccctgaag ctcgtgggcg ggggccacct gcactgcaga
     1321 ctggaaacca catacagatc caagaaattg agtgctaaag gtaagtgccc agtatgtgga
     1381 gcagagctga ctgacgaatt ctggagtcat ccacaattcg aaaagggtgg tggtagcggc
     1441 ggcggctccg gaggatcagc ttggtctcac cctcagtttg agaaataaaa gcttggctgt
     1501 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1561 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1621 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1681 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1741 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1801 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1861 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1921 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1981 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2041 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2101 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2161 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2221 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2281 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2341 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2401 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2461 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2521 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2581 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2641 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2701 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2761 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2821 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2881 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2941 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3001 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3061 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3121 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3181 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3241 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3301 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3361 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3421 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3481 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3541 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3601 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3661 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3721 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3781 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3841 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3901 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3961 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4021 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4081 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     4141 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     4201 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     4261 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     4321 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4381 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4441 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4501 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4561 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4621 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     4681 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     4741 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     4801 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     4861 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     4921 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     4981 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     5041 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     5101 acttttcata ctcccgccat tcagag
//
