LOCUS       sequence-479117-        8630 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  This plasmid encodes a genetically encoded fluorescent biosensor 
            based on human CXCR3 for chemokine CXCL9-11 detection. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8630)
  TITLE     Spatiotemporal mapping of multidimensional chemokine dynamics by 
            genetically encoded fluorescent sensors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pDisplay-CMV-GRAB_LoX3-1.0
FEATURES             Location/Qualifiers
     source          1..8630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     737..799
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1930..1951
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2057..2078)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2318..2337)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(2829..3402,3403..3405,3406..3414)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2996..3013)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3222..3241
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3415..4122
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3560..3578)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3828..3847
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4180..4209
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4213..4359
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(4381..4398)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4387..4611
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4743..5331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4823..4842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5660..5707
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5732..5751
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5939..6733)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTYGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6050..6069)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6660..6679
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(6832..6851)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        6930..7121
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7078..7098
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(7181..7972,7973..8041))
                     /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     7804..7823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8042..8146)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8173..8628
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8260..8279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8470..8491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccctgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
      721 cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
      781 tccaggttcc actggtgaca caagtttgta caaaaaagtt ggcatggtcc ttgaggtgag
      841 tgaccaccaa gtgctaaatg acgccgaggt tgccgccctc ctggagaact tcagctcttc
      901 ctatgactat ggagaaaacg agagtgactc gtgctgtacc tccccgccct gcccacagga
      961 cttcagcctg aacttcgacc gggccttcct gccagccctc tacagcctcc tctttctgct
     1021 ggggctgctg ggcaacggcg cggtggcagc cgtgctgctg agccggcgga cagccctgag
     1081 cagcaccgac accttcctgc tccacctagc tgtagcagac acgctgctgg tgctgacact
     1141 gccgctctgg gcagtggacg ctgccgtcca gtgggtcttt ggctctggcc tctgcaaagt
     1201 ggcaggtgcc ctcttcaata tcaacttcta cgcaggagcc ctcctgctgg cctgcatcag
     1261 ctttgaccgc tacctgaaca tagttcatgc cacccagctc taccgccggg ggcccccggc
     1321 ccgcgtgacc ctcacctgcc tggctgtctg ggggctctgc ctgcttttcg ccctcccaga
     1381 cttcatcttc ctgtcggccc accacgacga gcgcctcaac gccacccact gccaatacaa
     1441 cttcccacag gtgggccgca cggctctgcg ggtgctgcag ctggtggctg gctttctgct
     1501 gcccctgctg gtcatggcct actgctatgc ccacatcctg gccgccgtcg ccgcgccgcc
     1561 ggggggcacc gagcgcaggc ccaacggtct gggccccgag cgcagcgcgg gcccgggggg
     1621 cgcagaggcc gaaccgctgc ccacccagct caacggcgcc cctggcgagc ccgcgccggc
     1681 cgggccgcgc gacaccgacg cgctggacct ggaggaggga ggaaacgtct atatcaaggc
     1741 cgacaagcag aagaacggca tcaaggcgaa cttctggatc cgccacaaca tcgaggacgg
     1801 cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1861 gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa
     1921 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1981 cgagctgtac aagggcggta ccggagggag catggtgaga aagggcgagg agctgttcac
     2041 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2101 gtccggcgag ggtgagggcg atgccaccga gggcaagctg accctgaagt tcatctgcac
     2161 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2221 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2281 cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2341 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2401 cttcaaggag gacggcaaca tcctggggca caagctggag tacaaccccg gagcagcagc
     2461 acgctggcgc gggatgcggc tggtggtggt ggtcgtggtg gcctttgccc tctgctggac
     2521 cccctatcac ctggtggtgc tggtggacat cctcatggac ctgggcgctt tggcccgcaa
     2581 ctgtggccga gaaagcaggg tagacgtggc caagtcggtc acctcaggcc tgggctacat
     2641 gcactgctgc ctcaacccgc tgctctatgc ctttgtaggg gtcaagttcc gggagcggat
     2701 gtggatgctg ctcttgcgcc tgggctgccc caaccagaga gggctccaga ggcagccatc
     2761 gtcttcccgc cgggattcat cctggtctga gacctcagag gcctcctact cgggcttgta
     2821 agctagcgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     2881 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     2941 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3001 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3061 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     3121 aggtgcctct gcggccaacg ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     3181 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     3241 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     3301 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     3361 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg
     3421 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     3481 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     3541 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     3601 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     3661 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     3721 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     3781 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     3841 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     3901 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     3961 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     4021 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     4081 gagggccgcc actccaccgg cggcatggac gagctgtaca aggatggtaa aaagaagaaa
     4141 aagaagtcaa agacaaagtg tgtaattatg taagtcgacg aacaaaaact catctcagaa
     4201 gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg
     4261 ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc
     4321 cttatcatcc tcatcatgct ttggcagaag aagccacgtt aggcggccgc tcgagatcag
     4381 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4441 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4501 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4561 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4621 cggaaagaac cagtggcggt aatacggtta tccacagaat cgggggataa cgcaggaaag
     4681 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4741 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4801 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4861 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4921 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4981 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5041 aactatcgtc ttgagtccaa cccggtaaga cgcgacttat cgccactggc agcagccact
     5101 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5161 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5221 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5281 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5341 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5401 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5461 aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg
     5521 ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa
     5581 acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     5641 gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt
     5701 attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc
     5761 cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     5821 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     5881 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     5941 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     6001 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     6061 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     6121 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca taggtcacga
     6181 cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga
     6241 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     6301 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     6361 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     6421 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     6481 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     6541 ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     6601 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     6661 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     6721 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct
     6781 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccgg gcctcggcct
     6841 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     6901 ttaggggcgg aatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     6961 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     7021 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     7081 ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata
     7141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8161 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     8221 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     8281 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     8341 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     8401 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     8461 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     8521 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     8581 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttac
//
