LOCUS       sequence-218600-        6374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express feline Jam-A transiently in cell lines that do not 
            naturally express the receptor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  AUTHORS   Lu Z, Ledgerwood ED, Hinchman MM, Dick R, Parker JSL
  TITLE     Conserved Surface Residues on the Feline Calicivirus Capsid Are 
            Essential for Interaction with Its Receptor Feline Junctional 
            Adhesion Molecule A (fJAM-A).
  JOURNAL   J Virol. 2018 Mar 28;92(8):e00035-18. doi: 10.1128/JVI.00035-18. 
            Print 2018 Apr 15.
  PUBMED    29386293
REFERENCE   2  (bases 1 to 6374)
  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: 
            "10.1128/JVI.00035-18"; journalName: "J Virol"; date: "2018-03-28- 
            28"; volume: "92"; issue: "8"; pages: "e00035-18"
COMMENT     Sequence Label: pCI-Neo-JAM-A
FEATURES             Location/Qualifiers
     source          1..6374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2078..2199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2089..2108)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2143..2162
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2385..2840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2472..2491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2682..2703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2957..3314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2957..3148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2980..3000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3165..3300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3365..4159
                     /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(3419..3438)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4029..4048
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4223..4271
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4312..4331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4431..4453
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4491..4509)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4577..4681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4682..4750,4751..5542)
                     /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     complement(4900..4919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5713..6301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6202..6221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc atcgccatgg ggaccgaggc gagagccggg aggagacagc tgctcgtctt
     1141 cacgtcggtc gtcctgagct ccctggcgtt gggcaggggc gcagtgtata cttctgagcc
     1201 cgatgtcaga gttcctgagg acaaacccgc caagttgtcc tgctcctact cgggcttctc
     1261 caacccccgc gtggagtgga agttcgccca tggtgacatc accagcctcg tttgttataa
     1321 aaacaagatc acggcctcct atgcagaccg agtcaccttc tcgcacagtg gcatcacttt
     1381 ccattcggtg acccggaaag acacggggac gtacacttgc atggtctctg acgatggcgg
     1441 caacacgtac ggggaggtca gcgtccagct cactgtgctt gtgcctccat ccaagcccac
     1501 ggtccacatc ccctcctctg ccaccatcgg gagtcgggca gtgctgacct gctcagagaa
     1561 ggatggctcc cccccttctg agtactactg gttcaaggac ggggtccgga tgcctttgga
     1621 gcccaagggc aaccgcgcct tcagcaactc ttcctacagc ctgaatgaaa agacagggga
     1681 gctggtcttc gatcccgtgt cagcctggga tacgggcgag tacacctgtg aggcgcagaa
     1741 cgggtacggg atgcccatga ggtcagaggc tgtgcgcatg gaagccgcgg agttgaatgt
     1801 ggggggcatt gtggcagctg tccttgtcac actcattctc cttggattct tgattttggg
     1861 catctggttc gcctatagac gaggctactt tgacagaaca aagaaaggga cctcgagtaa
     1921 gaaggtgatt tacagccagc ccgctgcccg cagtgaaggg gaattcaggc agacctcgtc
     1981 attcctggtg tgaccctggt cccacgcgtg gtacctctag agtcgacccg ggcggccgct
     2041 tccctttagt gagggttaat gcttcgagca gacatgataa gatacattga tgagtttgga
     2101 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2161 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2221 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     2281 aaatgtggta aaatccgata aggatcgatc cgggctggcg taatagcgaa gaggcccgca
     2341 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     2401 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2461 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2521 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2581 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2641 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2701 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2761 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2821 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2881 tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     2941 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3061 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3121 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3181 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3241 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3301 taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc
     3361 caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3421 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3481 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3541 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3601 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3661 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3721 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3781 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3841 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3901 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3961 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4021 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4081 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4141 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4201 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     4261 ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca gtacaatctg
     4321 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4381 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4441 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4501 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4561 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4621 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4681 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4741 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4801 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4861 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4921 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4981 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5041 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5101 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5161 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5221 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5281 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5341 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5401 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5461 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5521 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5581 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5641 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5821 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6361 ggctcgacag atct
//
