LOCUS       sequence-353275-        6412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GPI-anchored dL5** in mammalian cells.  (FAP-GPI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6412)
  AUTHORS   Zhong Z, Ning J, Boggs EA, Jang S, Wallace C, Telmer C, Bruchez MP, 
            Ahn J, Engelman AN, Zhang P, Watkins SC, Ambrose Z
  TITLE     Cytoplasmic CPSF6 Regulates HIV-1 Capsid Trafficking and Infection 
            in a Cyclophilin A-Dependent Manner.
  JOURNAL   mBio. 2021 Mar 23;12(2). pii: mBio.03142-20. doi: 
            10.1128/mBio.03142-20.
  PUBMED    33758083
REFERENCE   2  (bases 1 to 6412)
  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: "mBio."; 
            date: "2021-03-23"; volume: "12(2). pii: mBio.03142-20. doi"; pages:
            " 10.1128/mBio.03142-20"
COMMENT     Sequence Label: pcDNA-IgKappa-myc-dL5- 2XG4S-GPI
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     949..1008
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1018..1047
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1063..1782
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     primer_bind     complement(2003..2020)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2009..2233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2279..2707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2366..2385)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2576..2597
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2716..2736)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2721..3050
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2901..3036
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2963..2982
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3117..3911
                     /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(3171..3190)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3781..3800
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4085..4206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4122..4141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4176..4195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4255..4271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4255..4271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4268..4290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4279..4295
                     /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(4303..4309,4310..4327,4328..4333))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4348..4369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4486..4503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4657..5245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4737..4756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5416..6207,6208..6276))
                     /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     6039..6058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6277..6381)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgc caccggatcc ggcttgggga tatccaccat ggagacagac
      961 acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga cgaattcgag
     1021 cagaagctga tcagcgagga ggacctgggc ctttctggcc tgcaggccgt cgttacccaa
     1081 gaacctagtg ttaccgttag cccaggtggt actgttatac ttacttgtgg aagtggtacg
     1141 ggtgccgtca catctggtca ttatgcaaat tggtttcaac aaaaaccagg acaagctcca
     1201 agagctttga tttttgatac tgataagaag tattcttgga ccccaggtag attttctgga
     1261 tctttgctgg gagcaaaggc agctttgaca atatcagatg ctcagcctga ggacgaagcc
     1321 gagtattact gttctcttag cgacgtggat ggctacttgt ttggcggtgg aacacaactg
     1381 acggttctgt ccggtggtgg cggctctggt ggcggtggca gcggcggtgg tggttccgga
     1441 ggcggcggtt ctcaggctgt ggtgactcag gagccgtcag tgactgtgtc cccaggaggg
     1501 acagtcattc tcacttgtgg ctccggcact ggagctgtca ccagtggtca ttatgccaac
     1561 tggttccagc agaagcctgg ccaagccccc agggcactta tatttgacac cgacaagaag
     1621 tattcctgga cccctggccg attctcaggc tccctccttg gggccaaggc tgccctgacc
     1681 atctcggatg cgcagcctga agatgaggct gagtattact gttcgctctc cgacgttgac
     1741 ggttatctgt tcggaggagg cacccagctg accgtcctct ccgcggccgg tctggcccat
     1801 atgggaggcg gtgggtctgg tggcggtggc agcaccggtc caaataaagg aagtggaacc
     1861 acatccggaa caacgcgact tctgtcaggg cacacgtgct tcacgctgac cggactgctt
     1921 ggaactttgg tgaccatggg cttgctgact tagctcgagg cagctgtcga ctctagaggg
     1981 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2041 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2101 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2161 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2221 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2281 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2341 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2401 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2461 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2521 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2581 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2641 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2701 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2821 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2881 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2941 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3001 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3061 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3121 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3181 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3241 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3301 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3361 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3421 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3481 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3541 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3601 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3661 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3721 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3781 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3841 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3901 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3961 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4021 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4081 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4141 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4201 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4261 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4321 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4441 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6361 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
