LOCUS       Exported                7103 bp ds-DNA     circular SYN 24-JUL-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7103)
  AUTHORS   Felix Horns
  TITLE     Direct Submission
  JOURNAL   Exported Jul 5, 2023 from SnapGene 4.0.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7103
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      923..932
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             929..1351
                     /codon_start=1
                     /product="Rev-independent version of HIV-1 Gag coding 
                     sequence from HXB2 [pCMV55M1-10 obtained from Dr. George 
                     Pavlakis, NCI (Schwartz et al (1992) J. Virol. 66: 
                     7176-7182]. "
                     /label=HIV-1 gag
                     /translation="MGARASVSPTSILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKN
                     WMTETLLVQNANPDCKTILKALGPAATLEEMMTACQGVGGPGHKARVLAEAMSQVTNSA
                     TIMLQRMKQLEDKVEELLSKNYHLENEVARLKKLVGE"
     CDS             932..949
                     /product="HIV-1 Matrix protein p17 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 MA
     CDS             950..1207
                     /label=HIV-1 CA
     CDS             1208..1249
                     /product="HIV-1 Spacer peptide 1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 SP1
     CDS             1250..1351
                     /product="Leucine zipper from GCN4. Sequence from 
                     https://jvi.asm.org/content/jvi/85/14/7419.full.pdf Fig 1B.
                     Original sequence from 
                     https://science.sciencemag.org/content/254/5031/539/tab-art
                     icle-info."
                     /label=GCN4 leucine zipper
     CDS             1352..1738
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
     CDS             1739..1894
                     /codon_start=1
                     /product="Rev-independent version of HIV-1 Gag coding 
                     sequence from HXB2 [pCMV55M1-10 obtained from Dr. George 
                     Pavlakis, NCI (Schwartz et al (1992) J. Virol. 66: 
                     7176-7182]. "
                     /label=HIV-1 gag
                     /translation="LQSRPEPTAPPEESFRSGVETTTPPQKQEPIDKELYPLTSLRSLF
                     GNDPSSQ"
     CDS             1739..1894
                     /product="Plays a role in budding of the assembled particle
                     by interacting with the host class E VPS proteins TSG101 
                     and PDCD6IP/AIP1."
                     /label=HIV-1 p6
     CDS             1757..1768
                     /product="Late-budding domains (L domains) are short 
                     sequence motifs essential for viral particle budding. They 
                     recruit proteins of the host ESCRT machinery (Endosomal 
                     Sorting Complex Required for Transport) or ESCRT-associated
                     proteins. p6-gag contains two L domains: a PTAP/PSAP motif,
                     which interacts with the UEV domain of TSG101 and a 
                     LYPX(n)L motif which interacts with PDCD6IP/AIP1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 p6-gag L domain
     CDS             1841..1870
                     /product="Late-budding domains (L domains) are short 
                     sequence motifs essential for viral particle budding. They 
                     recruit proteins of the host ESCRT machinery (Endosomal 
                     Sorting Complex Required for Transport) or ESCRT-associated
                     proteins. p6-gag contains two L domains: a PTAP/PSAP motif,
                     which interacts with the UEV domain of TSG101 and a 
                     LYPX(n)L motif which interacts with PDCD6IP/AIP1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 p6-gag L domain
     CDS             1895..1951
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1952..2668
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    2700..2924
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2970..3398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3412..3741
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3592..3727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3808..4602
                     /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"
     polyA_signal    4776..4897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4970..4986
                     /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(4994..5024)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5039..5060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5348..5936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6107..6967)
                     /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"
     promoter        complement(6968..7072)
                     /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 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgccaccat gggtgcgaga gcgtcagtaa gccctaccag
      961 cattctggac ataagacaag gaccaaaaga accctttaga gactatgtag accggttcta
     1021 taaaactcta agagctgagc aagcttcaca ggaggtaaaa aattggatga cagaaacctt
     1081 gttggtccaa aatgcgaacc cagattgtaa gaccatcctg aaggctctcg gcccagcggc
     1141 tacactagaa gaaatgatga cagcatgtca gggagtagga ggacccggcc ataaggcaag
     1201 agttttggct gaagcaatga gccaagtaac aaattcagct accataatgc tccaaagaat
     1261 gaaacagctt gaggataagg tcgaggaact cctctccaaa aactaccatc tggagaacga
     1321 agtggctcgg ctcaagaaac ttgtgggtga agcttcaaac tttactcagt tcgtgctcgt
     1381 ggacaatggt gggacagggg atgtgacagt ggctccttct aatttcgcta atggggtggc
     1441 agagtggatc agctccaact cacggagcca ggcctacaag gtgacatgca gcgtcaggca
     1501 gtctagtgcc cagaagagaa agtataccat caaggtggag gtccccaaag tggctaccca
     1561 gacagtgggc ggagtcgaac tgcctgtcgc cgcttggagg tcctacctga acatggagct
     1621 cactatccca attttcgcta ccaattctga ctgtgaactc atcgtgaagg caatgcaggg
     1681 gctcctcaaa gacggtaatc ctatcccttc cgccatcgcc gctaactcag gtatctacct
     1741 tcagagcaga ccagagccaa cagccccacc agaagagagc ttcaggtctg gggtagagac
     1801 aacaactccc cctcagaagc aggagccgat agacaaggaa ctgtatcctt taacttccct
     1861 cagatcactc tttggcaacg acccctcgtc acaagctact aacttcagcc tgctgaagca
     1921 ggctggagac gtggaggaga accctggacc tatgagcaag ggcgaggagc tgttcaccgg
     1981 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2041 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2101 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2161 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2221 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2281 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2341 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2401 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2461 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2521 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2581 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2641 tcacggcatg gacgagctgt acaagtaagg gcccgtttaa acccgctgat cagcctcgac
     2701 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2761 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2821 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2881 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2941 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3001 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3061 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3121 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3181 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3241 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3301 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3361 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3421 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3541 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3601 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3661 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3721 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3781 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3841 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3901 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3961 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4021 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4081 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4141 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4201 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4261 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4321 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4381 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4441 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4501 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4561 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4621 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4681 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4741 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4801 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4861 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4921 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4981 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5041 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5101 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5161 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5221 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5281 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5341 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5401 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5461 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5521 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5581 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5641 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5701 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5761 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5821 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5881 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5941 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6001 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6061 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6241 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7081 cccgaaaagt gccacctgac gtc
//
