LOCUS       Exported                7757 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 7757)
  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..7757
                     /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..2161
                     /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="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
                     IAPGQMREPRGSDIAGTTSTLQEQIGWMTNNPPIPVGEIYKRWIILGLNKIVRMYSPTS
                     ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPA
                     ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNSATIMLQRMKQLEDKVEELLSKNYHLE
                     NEVARLKKLVGE"
     CDS             932..1324
                     /product="HIV-1 Matrix protein p17 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 MA
     CDS             1325..2017
                     /label=HIV-1 CA
     CDS             2018..2059
                     /product="HIV-1 Spacer peptide 1 
                     (https://www.uniprot.org/uniprot/P04591)"
                     /label=HIV-1 SP1
     CDS             2060..2161
                     /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             2162..2548
                     /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             2549..2605
                     /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             2606..3322
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    3354..3578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3624..4052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4066..4395
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4246..4381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4462..5256
                     /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    5430..5551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5600..5616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5624..5640
                     /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(5648..5678)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5693..5714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6002..6590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6761..7621)
                     /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(7622..7726)
                     /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 gcgtcagtat taagcggggg
      961 agaattagat cgatgggaaa aaattcggtt aaggccaggg ggaaagaaga agtacaagct
     1021 aaagcacatc gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     1081 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     1141 atcagaggag cttcgatcac tatacaacac agtagcaacc ctctattgtg tgcaccagcg
     1201 gatcgagatc aaggacacca aggaagcttt agacaagata gaggaagagc aaaacaagtc
     1261 caagaagaag gcccagcagg cagcagctga cacaggacac agcaatcagg tcagccaaaa
     1321 ttaccctata gtgcagaaca tccaggggca aatggtacat caggccatat cacctagaac
     1381 tttaaatgca tgggtaaaag tagtagaaga gaaggctttc agcccagaag tgatacccat
     1441 gttttcagca ttatcagaag gagccacccc acaggacctg aacacgatgt tgaacaccgt
     1501 ggggggacat caagcagcca tgcaaatgtt aaaagagacc atcaatgagg aagctgcaga
     1561 atgggataga gtgcatccag tgcatgcagg gcctattgca ccaggccaga tgagagaacc
     1621 aaggggaagt gacatagcag gaactactag tacccttcag gaacaaatag gatggatgac
     1681 aaataatcca cctatcccag taggagagat ctacaagagg tggataatcc tgggattgaa
     1741 caagatcgtg aggatgtata gccctaccag cattctggac ataagacaag gaccaaaaga
     1801 accctttaga gactatgtag accggttcta taaaactcta agagctgagc aagcttcaca
     1861 ggaggtaaaa aattggatga cagaaacctt gttggtccaa aatgcgaacc cagattgtaa
     1921 gaccatcctg aaggctctcg gcccagcggc tacactagaa gaaatgatga cagcatgtca
     1981 gggagtagga ggacccggcc ataaggcaag agttttggct gaagcaatga gccaagtaac
     2041 aaattcagct accataatgc tccaaagaat gaaacagctt gaggataagg tcgaggaact
     2101 cctctccaaa aactaccatc tggagaacga agtggctcgg ctcaagaaac ttgtgggtga
     2161 agcttcaaac tttactcagt tcgtgctcgt ggacaatggt gggacagggg atgtgacagt
     2221 ggctccttct aatttcgcta atggggtggc agagtggatc agctccaact cacggagcca
     2281 ggcctacaag gtgacatgca gcgtcaggca gtctagtgcc cagaagagaa agtataccat
     2341 caaggtggag gtccccaaag tggctaccca gacagtgggc ggagtcgaac tgcctgtcgc
     2401 cgcttggagg tcctacctga acatggagct cactatccca attttcgcta ccaattctga
     2461 ctgtgaactc atcgtgaagg caatgcaggg gctcctcaaa gacggtaatc ctatcccttc
     2521 cgccatcgcc gctaactcag gtatctacgc tactaacttc agcctgctga agcaggctgg
     2581 agacgtggag gagaaccctg gacctatgag caagggcgag gagctgttca ccggggtggt
     2641 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2701 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2761 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2821 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2881 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2941 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3001 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3061 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3121 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3181 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3241 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactcacgg
     3301 catggacgag ctgtacaagt aagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3541 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3601 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3661 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3721 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3781 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3841 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3901 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3961 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4021 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4081 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4141 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4201 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4261 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4321 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4381 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4441 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4501 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4561 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4621 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4681 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4741 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4801 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4861 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4921 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4981 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5041 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5101 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5161 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5221 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5281 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5341 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5401 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5461 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5521 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5581 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5641 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5701 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5761 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6181 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6421 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7741 aagtgccacc tgacgtc
//
