LOCUS       sequence-152867-        6444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of nonconjugative ISG15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6444)
  AUTHORS   Gonzalez-Sanz R, Mata M, Bermejo-Martin J, Alvarez A, Cortijo J, 
            Melero JA, Martinez I
  TITLE     ISG15 Is Upregulated in Respiratory Syncytial Virus Infection and 
            Reduces Virus Growth through Protein ISGylation.
  JOURNAL   J Virol. 2016 Jan 13;90(7):3428-38. doi: 10.1128/JVI.02695-15.
  PUBMED    26763998
REFERENCE   2  (bases 1 to 6444)
  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"; 
            journalName: "J Virol"; date: "2016-01-13- 13"; volume: "90"; issue:
            "7"; pages: "3428-38"
COMMENT     Sequence Label: pCMV6-Neo-ISG15-LRAA
FEATURES             Location/Qualifiers
     source          1..6444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1641..1657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1668..2290
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1867..1886)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2314..2334)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2319..2648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2499..2634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2715..3509
                     /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(2769..2788)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3379..3398
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3683..3804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3720..3739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3774..3793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3853..3869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3866..3888)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3877..3893
                     /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(3901..3907,3908..3925,3926..3931))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3946..3967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4084..4101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4255..4840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4335..4354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5011..5802,5803..5871))
                     /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     5634..5653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5872..5976)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6003..6431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6090..6109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6300..6321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccgcgg tgctgcctgc cgaagccggc ggctgagagg cagcgaactc
     1021 atctttgcca gtacaggagc tcgtgccgtg gcccacagcc cacagcccac agccatgggc
     1081 tgggacctga cggtgaagat gctggcgggc aacgaattcc aggtgtccct gagcagctcc
     1141 atgtcggtgt cagagctgaa ggcgcagatc acccagaaga tcggcgtgca cgccttccag
     1201 cagcgtctgg ctgtccaccc gagcggtgtg gcgctgcagg acagggtccc ccttgccagc
     1261 cagggcctgg gccccggcag cacggtcctg ctggtggtgg acaaatgcga cgaacctctg
     1321 agcatcctgg tgaggaataa caagggccgc agcagcacct acgaggtgcg gctgacgcag
     1381 accgtggccc acctgaagca gcaagtgagc gggctggagg gtgtgcagga cgacctgttc
     1441 tggctgacct tcgaggggaa gcccctggag gaccagctcc cgctggggga gtacggcctc
     1501 aagcccctga gcaccgtgtt catgaatctg cgcctgcggg cagccggcac agagcctggc
     1561 gggcggagct aagggcctcc accagcatcc gagcaggatc aagggccgga aataaaggct
     1621 gttgtaaaga ggcggccgcg gtcatagctg tttcctgaac agatcccggg tggcatccct
     1681 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     1741 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     1801 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     1861 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     1921 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     1981 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     2041 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     2101 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     2161 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     2221 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     2281 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     2341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2401 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2461 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2521 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2581 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2641 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     2701 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2761 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2821 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2881 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2941 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3001 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3061 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3121 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3181 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3241 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3301 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3361 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3421 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3481 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3541 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3601 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3661 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     3721 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3781 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     3841 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3901 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3961 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4021 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     4801 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4921 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4981 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5041 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5101 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5161 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5221 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5281 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5341 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5401 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5461 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5521 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5581 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5641 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5701 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5761 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5821 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5881 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5941 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6001 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6061 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6121 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6181 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     6241 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6301 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6361 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6421 taacgcgaat tttaacaaaa tatt
//
