LOCUS       Exported                9383 bp ds-DNA     circular SYN 03-FEB-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pL-GW-EcoDam-V5-mseEMD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9383)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Feb 3, 2022 from SnapGene 5.0.4
            https://www.snapgene.com
COMMENT     Generated by Lars Guelen
            ORIGDB|GenBank
FEATURES             Location/Qualifiers
     source          1..9383
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1056..1181
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1674..1907
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2092..2136
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2398..2878
                     /label=HSP
                     /note="/vntifkey=29"
     promoter        2598..2836
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     misc_feature    2885..2890
                     /label=Kozak
                     /note="/vntifkey=21"
     misc_feature    2891..3724
                     /label=EcoDam
                     /note="/vntifkey=21"
     misc_feature    3731..3772
                     /label=V5-tag
                     /note="/vntifkey=21"
     CDS             3731..3772
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    3788..3812
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3833..4612
                     /label=Emerin(Mm)
     protein_bind    complement(4629..4653)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     LTR             5184..5364
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5396..5620
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5666..6094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6108..6438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6289..6424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6486..6533
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6552..6926
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7056..7177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7226..7242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7250..7266
                     /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(7274..7304)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7319..7340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7628..8216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8387..9247)
                     /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(9248..9352)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cctgaaagcg aaagggaaac cagagctctc tcgacgcagg actcggcttg
     1081 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1141 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1201 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1261 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1321 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1381 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1441 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1501 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1561 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1621 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1681 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcctc aatgacgctg
     1741 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1801 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1861 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1921 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1981 tctctggaac agatctggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     2041 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     2101 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     2161 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2221 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     2281 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     2341 ggagagagag acagagacag atccattcga ttagtgaacg gatcccccgg gctgcaggaa
     2401 ttcagatctc ggccgcatat taagtgcatt gttctcgata ccgctaagtg cattgttctc
     2461 gttagctcga tggacaagtg cattgttctc ttgctgaaag ctcgatggac aagtgcattg
     2521 ttctcttgct gaaagctcga tggacaagtg cattgttctc ttgctgaaag ctcagtaccc
     2581 gggagtaccc tcgaccgccg gagtataaat agaggcgctt cgtctacgga gcgacaattc
     2641 aattcaaaca agcaaagtga acacgtcgct aagcgaaagc taagcaaata aacaagcgca
     2701 gctgaacaag ctaaacaatc tgcagtaaag tgcaagttaa agtgaatcaa ttaaaagtaa
     2761 ccagcaacca agtaaatcaa ctgcaactac tgaaatctgc caagaagtaa ttattgaata
     2821 caagaagaga actctgaata ctttcaacaa gttaccgaga aagaagaact cacacacagc
     2881 tagcgccacc atgaagaaaa atcgcgcttt tttgaagtgg gcagggggca agtatcccct
     2941 gcttgatgat attaaacggc atttgcccaa gggcgaatgt ctggttgagc cttttgtagg
     3001 tgccgggtcg gtgtttctca acaccgactt ttctcgttat atccttgccg atatcaatag
     3061 cgacctgatc agtctctata acattgtgaa gatgcgtact gatgagtacg tacaggccgc
     3121 acgcgagctg tttgttcccg aaacaaattg cgccgaggtt tactatcagt tccgcgaaga
     3181 gttcaacaaa agccaggatc cgttccgtcg ggcggtactg tttttatatt tgaaccgcta
     3241 cggttacaac ggcctgtgtc gttacaatct gcgcggtgag tttaacgtgc cgttcggccg
     3301 ctacaaaaaa ccctatttcc cggaagcaga gttgtatcac ttcgctgaaa aagcgcagaa
     3361 tgcctttttc tattgtgagt cttacgccga tagcatggcg cgcgcagatg atgcatccgt
     3421 cgtctattgc gatccgcctt atgcaccgct gtctgcgacc gccaacttta cggcgtatca
     3481 cacaaacagt tttacgcttg aacaacaagc gcatctggcg gagatcgccg aaggtctggt
     3541 tgagcgccat attccagtgc tgatctccaa tcacgatacg atgttaacgc gtgagtggta
     3601 tcagcgcgca aaattgcatg tcgtcaaagt tcgacgcagt ataagcagca acggcggcac
     3661 acgtaaaaag gtggacgaac tgctggcttt gtacaaacca ggagtcgttt cacccgcgaa
     3721 aaaaaccggt ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgatcgattt
     3781 cgatcaccca agtttgtaca aaaaagcagg ctccgcggcc gcccccttca ccatggacga
     3841 ctatgcggtt ttgtccgata ctgagctggc cgcagtgcta cgccagtaca acatccctca
     3901 tgggcctatt gtgggttcca ctcgcaagct ctacgaaaag aaaatcttcg agtacgagac
     3961 ccagagaagg agactcttac cccccaactc gtcatcttct tcattctcct atcagttctc
     4021 agacttggat tcagccgccg tggactcaga catgtatgat ctgcccaaaa aagaagacgc
     4081 cttactttac cagagcaagg actataatga tgactactat gaggagagtt atttgaccac
     4141 caagacatac ggggagcctg agtctgtggg catgtccaag agcttccgcc agccggggac
     4201 ctcacttgta gatgctgata ccttccatca ccaggtgcgt gatgacattt tctcttcttt
     4261 agaagaggaa ggcaaggata gggagcgcct catctatggc caagacagtg cctaccagag
     4321 catcgcacac taccgcccca tttccaatgt ctccaggagc tctctgggct tatcatatta
     4381 tcctacatcc tccacctctt ctgtgtcctc ctcttcatct tctccctctt catggcttac
     4441 ccgacgtgcc attcggccag agaagcaggc ccctgctgct gctctgggcc aggatcgaca
     4501 ggtcccactc tggggtcagc tgttactctt cctggtcttt gctgcctttc tgctctttgt
     4561 ttactattcc atacaagctg aagaaggcaa ccccttctgg atggatccct aaaagggtgg
     4621 gcgcgccgac ccagctttct tgtacaaagt ggttcgatcg aaccgcggtc tagactcgag
     4681 acctggaaaa acatggagca atcacaagta gcaacacagc agctaccaat gctgcttgtg
     4741 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     4801 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     4861 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     4921 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
     4981 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
     5041 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
     5101 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
     5161 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5221 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5281 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5341 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
     5401 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     5461 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     5521 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     5581 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     5641 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5701 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5761 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5821 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5881 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5941 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6001 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     6061 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     6121 ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat
     6181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     6301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6361 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6421 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     6481 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     6541 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     6601 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     6661 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     6721 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     6781 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     6841 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     6901 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     6961 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     7021 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     7081 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7141 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     7201 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7261 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7321 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7381 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7801 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     7861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8041 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     8101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9361 cccgaaaagt gccacctgac gtc
//
