LOCUS       sequence-210317-        5811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for human CCR5 coding sequence tagged with the mTFP1 FP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5811)
  AUTHORS   Iliopoulou M, Nolan R, Alvarez L, Watanabe Y, Coomer CA, 
            Jakobsdottir GM, Bowden TA, Padilla-Parra S
  TITLE     A dynamic three-step mechanism drives the HIV-1 pre-fusion reaction.
  JOURNAL   Nat Struct Mol Biol. 2018 Sep;25(9):814-822. doi: 
            10.1038/s41594-018-0113-x. Epub 2018 Aug 27.
  PUBMED    30150645
REFERENCE   2  (bases 1 to 5811)
  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: 
            "10.1038/s41594-018-0113-x"; journalName: "Nat Struct Mol Biol"; 
            date: "2018-09"; volume: "25"; issue: "9"; pages: "814-822"
COMMENT     Sequence Label: hCCR5-mTFP1
FEATURES             Location/Qualifiers
     source          1..5811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        104..407
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..611
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     562..582
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    821..832
                     /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      1739..1748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1745..2452
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     misc_feature    2453..2518
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2642..2763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2733..2752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2770..3225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2907..2928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3119..3138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3252..3356
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3358..3715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3358..3549
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3381..3401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3566..3701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3750..4544
                     /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             3750..4544
                     /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(3804..3823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4414..4433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4732..4751)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4776..4823
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5152..5740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5641..5660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
       61 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      121 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      181 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      241 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      301 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      361 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      421 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      481 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      541 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      601 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
      661 gctcaagctt atggattatc aagtgtcaag tccaatctat gacatcaatt attatacatc
      721 ggagccctgc caaaaaatca atgtgaagca aatcgcagcc cgcctcctgc ctccgctcta
      781 ctcactggtg ttcatctttg gttttgtggg caacatgctg gtcatcctca tcctgataaa
      841 ctgcaaaagg ctgaagagca tgactgacat ctacctgctc aacctggcca tctctgacct
      901 gtttttcctt cttactgtcc ccttctgggc tcactatgct gccgcccagt gggactttgg
      961 aaatacaatg tgtcaactct tgacagggct ctattttata ggcttcttct ctggaatctt
     1021 cttcatcatc ctcctgacaa tcgataggta cctggctgtc gtccatgctg tgtttgcttt
     1081 aaaagccagg acggtcacct ttggggtggt gacaagtgtg atcacttggg tggtggctgt
     1141 gtttgcgtct ctcccaggaa tcatctttac cagatctcaa aaagaaggtc ttcattacac
     1201 ctgcagctct cattttccat acagtcagta tcaattctgg aagaatttcc agacattaaa
     1261 gatagtcatc ttggggctgg tcctgccgct gcttgtcatg gtcatctgct actcgggaat
     1321 cctaaaaact ctgcttcggt gtcgaaatga gaagaagagg cacagggctg tgaggcttat
     1381 cttcaccatc atgattgttt attttctctt ctgggctccc tacaacattg tccttctcct
     1441 gaacaccttc caggaattct ttggcctgaa taattgcagt agctctaaca ggttggacca
     1501 agctatgcag gtgacagaga ctcttgggat gacgcactgc tgcatcaacc ccatcatcta
     1561 tgcctttgtc ggggagaagt tcagaaacta cctcttagtc ttcttccaaa agcacattgc
     1621 caaacgcttc tgcaaatgct gttctatttt ccagcaagag gctcccgagc gagcaagctc
     1681 agtttacacc cgatccactg gggagcagga aatatctgtg ggcttgttat taccggtcgc
     1741 caccatggtg agcaagggcg aggagaccac aatgggcgta atcaagcccg acatgaagat
     1801 caagctgaag atggagggca acgtgaatgg ccacgccttc gtgatcgagg gcgagggcga
     1861 gggcaagccc tacgacggca ccaacaccat caacctggag gtgaaggagg gagcccccct
     1921 gcccttctcc tacgacattc tgaccaccgc gttcgcctac ggcaacaggg ccttcaccaa
     1981 gtaccccgac gacatcccca actacttcaa gcagtccttc cccgagggct actcttggga
     2041 gcgcaccatg accttcgagg acaagggcat cgtgaaggtg aagtccgaca tctccatgga
     2101 ggaggactcc ttcatctacg agatacacct caagggcgag aacttccccc ccaacggccc
     2161 cgtgatgcag aagaagacca ccggctggga cgcctccacc gagaggatgt acgtgcgcga
     2221 cggcgtgctg aagggcgacg tcaagcacaa gctgctgctg gagggcggcg gccaccaccg
     2281 cgttgacttc aagaccatct acagggccaa gaaggcggtg aagctgcccg actatcactt
     2341 tgtggaccac cgcatcgaga tcctgaacca cgacaaggac tacaacaagg tgaccgttta
     2401 cgagagcgcc gtggcccgca attccaccga cggcatggac gagctgtaca agtccggact
     2461 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac
     2521 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2581 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2641 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2761 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2821 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2881 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2941 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3001 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3061 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3121 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3181 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3361 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3721 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3781 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3841 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3901 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3961 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4021 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4081 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4141 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4201 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4261 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4321 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4381 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4441 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4501 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4561 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4621 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4681 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4741 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4801 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4861 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4921 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4981 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5341 tgtatcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg c
//
