LOCUS       sequence-479118-        8629 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  This plasmid encodes a mutated version of the genetically encoded 
            chemokine biosensor LoX3-1.0, which is designed to be non-responsive
            to chemokine binding..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8629)
  TITLE     Spatiotemporal mapping of multidimensional chemokine dynamics by 
            genetically encoded fluorescent sensors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8629)
  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            "Unpublished"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-mut
FEATURES             Location/Qualifiers
     source          1..8629
                     /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..5330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4822..4841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5659..5706
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5731..5750
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5938..6732)
                     /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(6049..6068)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6659..6678
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(6831..6850)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        6929..7120
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7077..7097
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(7180..7971,7972..8040))
                     /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     7803..7822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8041..8145)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8172..8627
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8259..8278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8469..8490
                     /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 acccactact 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 gccgctccag gcagtgaaag 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 aatacggcta tccacagaat cgggggataa cgcaggaaag
     4681 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4741 tttttccata ggctccgccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4801 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4861 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4921 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4981 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5041 actatcgtct tgagtccaac ccggtaagac gcgacttatc gccactggca gcagccactg
     5101 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5161 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5221 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5401 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5461 aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt
     5521 tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa
     5581 cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg
     5641 accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta
     5701 ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc
     5761 ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     5821 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     5881 ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca
     5941 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     6001 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     6061 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     6121 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat aggtcacgac
     6181 gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag
     6241 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     6301 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     6361 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     6421 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     6481 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     6541 tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     6601 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     6661 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     6721 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta
     6781 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccggg cctcggcctc
     6841 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     6901 taggggcgga atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     6961 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     7021 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     7081 taactgacac acattccaca gctggttctt tccgcctcag gactcttcct ttttcaataa
     7141 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7261 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7321 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7381 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7441 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7501 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7561 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7621 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7681 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7741 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7801 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7861 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7921 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7981 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8041 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8101 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8161 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     8221 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     8281 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     8341 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     8401 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     8461 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     8521 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     8581 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttac
//
