LOCUS       sequence-169209-        7128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ecliptic pHluorin–ICAM-1 (intercellular adhesion molecule 
            1) in mammalian cells. Incorporates into viral membrane to label 
            pseudoviruses.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7128)
  AUTHORS   Miyauchi K, Marin M, Melikyan GB
  TITLE     Visualization of retrovirus uptake and delivery into acidic 
            endosomes.
  JOURNAL   Biochem J. 2011 Mar 15;434(3):559-69. doi: 10.1042/BJ20101588.
  PUBMED    21175427
REFERENCE   2  (bases 1 to 7128)
  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"; journalName: 
            "Biochem J"; date: "2011-03-15- 15"; volume: "434"; issue: "3"; 
            pages: "559-69"
COMMENT     Sequence Label: EcpH-TM
FEATURES             Location/Qualifiers
     source          1..7128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             932..976
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             977..1042
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1052..1762
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1079..1107)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1702..1722
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    1869..2491
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2068..2087)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2515..2535)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2533..2831
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2885..3679
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2939..2958)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3549..3568
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4331..4465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4368..4387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4422..4441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4470..4489)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4471..4489)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4503..4519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4503..4519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4516..4538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4527..4543
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4551..4557,4558..4575,4576..4581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4596..4617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4734..4751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4905..5493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4985..5004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5664..6455,6456..6524))
                     /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     6287..6306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6525..6629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6743..6762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6970..6991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcac catgtctgca cttctgatcc tagctcttgt
      961 tggagctgca gttgctgact acaaagacca tgacggtgat tataaagatc atgacatcga
     1021 ttacaaggat gacgatgaca agcttctcga gagtaaagga gaagaacttt tcactggagt
     1081 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     1141 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     1201 aaaactacct gttccttggc caacacttgt cactacttta acttatggtg ttcaatgctt
     1261 ttcaagatac ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg
     1321 ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga
     1381 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     1441 agaagatgga aacattcttg gacacaaatt ggaatacaac tataacgatc accaggtgta
     1501 catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta gacacaacat
     1561 tgaagatgga ggcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     1621 gcccgtcctt ttaccagaca accattacct gtttacaact tctactcttt cgaaagatcc
     1681 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     1741 tggcatggat gaactataca agaattcacg gtatgagatt gtcatcatca ctgtggtagc
     1801 agccgcagtc ataatgggca ctgcaggcct cagcacgtac ctctataacc gccagcggtg
     1861 aggatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     1921 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     1981 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2041 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2101 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2161 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2221 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2281 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     2341 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     2401 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     2461 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg
     2521 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2581 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2641 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2701 tcccgcctcc gctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2761 tttttattta tgcaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2821 ggcttttttg gaggaattga tcagcttggg atctgatcaa gagacaggat gaggatcgtt
     2881 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2941 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3001 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3061 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3121 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3181 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3241 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3301 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3361 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3421 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3481 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3541 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3601 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3661 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3721 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3781 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3841 ttcgcccacc ccgggctcga tcccctcgcg agttggttca gctgctgcct gaggctggac
     3901 gacctcgcgg agttctaccg gcagtgcaaa tccgtcggca tccaggaaac cagcagcggc
     3961 tatccgcgca tccatgcccc cgaactgcag gagtggggag gcacgatggc cgctttggtc
     4021 gacccggacg ggacgctcct gcgcctgata cagaacgaat tgcttgcagg catctcatga
     4081 gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat ggagcgctgg cgcctgctgc
     4141 gcgacggcga gctgctcacc acccactcgc caagctggaa ccgtaaaaag gccgcgttgc
     4201 tggcgttttt ccataggctc cgccgatcat aatcagccat accacatttg tagaggtttt
     4261 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4321 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4381 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4441 caatgtatct tatcatgtct ggatcaattc cctatagtga gtcgtattaa attcgtaatc
     4501 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5341 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6541 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6601 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     6661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa gagtccacta
     6961 ttaaagaacg tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     7021 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     7081 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaattt
//
