LOCUS       sequence-210324-        5807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for human CCR5 coding sequence tagged with the LSS-mOrange 
            FP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5807)
  AUTHORS   Starling T, Carlon-Andres I, Iliopoulou M, Kraemer B, 
            Loidolt-Krueger M, Williamson DJ, Padilla-Parra S
  TITLE     Multicolor lifetime imaging and its application to HIV-1 uptake.
  JOURNAL   Nat Commun. 2023 Aug 17;14(1):4994. doi: 10.1038/s41467-023-40731-x.
  PUBMED    37591879
REFERENCE   2  (bases 1 to 5807)
  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: "Nat 
            Commun."; date: "2023-08-17"; volume: "14(1):4994. doi"; pages: " 
            10.1038/s41467-023-40731-x"
COMMENT     Sequence Label: hCCR5-LSSmOrange
FEATURES             Location/Qualifiers
     source          1..5807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        411..614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    824..835
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     regulatory      1742..1751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1748..2455
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2161..2180
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2461..2517
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2641..2762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2678..2697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2732..2751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2769..3224)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2906..2927)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3118..3137
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3251..3355
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3357..3714
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3357..3548
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3380..3400)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3565..3700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3749..4543
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3749..4543
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3803..3822)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4413..4432
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4731..4750)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4775..4822
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5151..5739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5640..5659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      661 cgagctcaag cttatggatt atcaagtgtc aagtccaatc tatgacatca attattatac
      721 atcggagccc tgccaaaaaa tcaatgtgaa gcaaatcgca gcccgcctcc tgcctccgct
      781 ctactcactg gtgttcatct ttggttttgt gggcaacatg ctggtcatcc tcatcctgat
      841 aaactgcaaa aggctgaaga gcatgactga catctacctg ctcaacctgg ccatctctga
      901 cctgtttttc cttcttactg tccccttctg ggctcactat gctgccgccc agtgggactt
      961 tggaaataca atgtgtcaac tcttgacagg gctctatttt ataggcttct tctctggaat
     1021 cttcttcatc atcctcctga caatcgatag gtacctggct gtcgtccatg ctgtgtttgc
     1081 tttaaaagcc aggacggtca cctttggggt ggtgacaagt gtgatcactt gggtggtggc
     1141 tgtgtttgcg tctctcccag gaatcatctt taccagatct caaaaagaag gtcttcatta
     1201 cacctgcagc tctcattttc catacagtca gtatcaattc tggaagaatt tccagacatt
     1261 aaagatagtc atcttggggc tggtcctgcc gctgcttgtc atggtcatct gctactcggg
     1321 aatcctaaaa actctgcttc ggtgtcgaaa tgagaagaag aggcacaggg ctgtgaggct
     1381 tatcttcacc atcatgattg tttattttct cttctgggct ccctacaaca ttgtccttct
     1441 cctgaacacc ttccaggaat tctttggcct gaataattgc agtagctcta acaggttgga
     1501 ccaagctatg caggtgacag agactcttgg gatgacgcac tgctgcatca accccatcat
     1561 ctatgccttt gtcggggaga agttcagaaa ctacctctta gtcttcttcc aaaagcacat
     1621 tgccaaacgc ttctgcaaat gctgttctat tttccagcaa gaggctcccg agcgagcaag
     1681 ctcagtttac acccgatcca ctggggagca ggaaatatct gtgggcttgt tattaccggt
     1741 cgccaccatg gtgagcaagg gcgaggagaa taacatggcc atcatcaagg agttcatgcg
     1801 cttcaaggtg cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1861 cgagggccgc ccctacgagg gctttcagac cgttaagctg aaggtgacca agggtggccc
     1921 cctgcccttc gcctgggaca tcttgtcccc tcagttcacc tacggctcca aggcctacgt
     1981 gaagcacccc gccgacatcc ccgactacct caagctgtcc ttccccgagg gcttcaagtg
     2041 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgactcagg actcctccct
     2101 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2161 ccccgtaatg cagaagaaga ccatgggcat ggaggcctcc tccgagcgga tgtaccccga
     2221 ggacggcgcc ctgaagggcg aggacaagct caggctgaag ctgaaggacg gcggccacta
     2281 cacctccgag gtcaagacca cctacaaggc caagaagccc gtgcagttgc ccggcgccta
     2341 catcgtcgac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     2401 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtactc
     2461 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc
     2521 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2581 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2641 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2701 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2761 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2821 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2881 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2941 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3001 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3061 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3121 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3181 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3241 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3301 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3361 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3421 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3481 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3541 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3601 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3661 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3721 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3781 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3841 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3901 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3961 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4021 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4081 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4141 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4201 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4261 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4321 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4381 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4441 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4501 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4561 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4621 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4681 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4741 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4801 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4861 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4921 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4981 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5161 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5221 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5281 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5521 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5581 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5701 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5761 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcc
//
