LOCUS       Exported                9327 bp DNA     circular SYN 05-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9327)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 5, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2711..2720
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2720..2743
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2756..2782
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2789..3109
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     protein_bind    3116..3140
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4458..4482)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4512..5063
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             5075..5674
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5717..6305
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6188..6199)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6380..6613
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6818..6922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6923..7783
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7954..8542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8953..8974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8989..9019
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9027..9043
                     /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)."
     primer_bind     9051..9067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcatt
     2221 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     2281 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     2341 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     2401 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     2461 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     2521 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc aggggcgggg
     2581 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     2641 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcggccgc agatctctcg
     2701 aggttgatct gccaccatgg actacaagga tgacgatgac aagctcgatg gaggataccc
     2761 atacgatgtt ccagattacg ctgctagcgg agtgcaggtg gaaaccatct ccccaggaga
     2821 cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg cactacaccg ggatgcttga
     2881 agatggaaag aaagttgatt cctcccggga cagaaacaag ccctttaagt ttatgctagg
     2941 caagcaggag gtgatccgag gctgggaaga aggggttgcc cagatgagtg tgggtcagag
     3001 agccaaactg actatatctc cagattatgc ctatggtgcc actgggcacc caggcatcat
     3061 cccaccacat gccactctcg tcttcgatgt ggagcttcta aaactggaaa cgcgtacaag
     3121 tttgtacaaa aaagcaggct tcaccatgag ctcccctggc accgagagcg cgggaaagag
     3181 cctgcagtac cgagtggacc acctgctgag cgccgtggag aatgagctgc aggcgggcag
     3241 cgagaagggc gaccccacag agcgcgaact gcgcgtgggc ctggaggaga gcgagctgtg
     3301 gctgcgcttc aaggagctca ccaatgagat gatcgtgacc aagaacggca ggaggatgtt
     3361 tccggtgctg aaggtgaacg tgtctggcct ggaccccaac gccatgtact ccttcctgct
     3421 ggacttcgtg gcggcggaca accaccgctg gaagtacgtg aacggggaat gggtgccggg
     3481 gggcaagccg gagccgcagg cgcccagctg cgtctacatc caccccgact cgcccaactt
     3541 cggggcccac tggatgaagg ctcccgtctc cttcagcaaa gtcaagctca ccaacaagct
     3601 caacggaggg ggccagatca tgctgaactc cttgcataag tatgagcctc gaatccacat
     3661 agtgagagtt ggggatccac agcgcatgat caccagccac tgcttccctg agacccagtt
     3721 catagtggtg actgcttatc agaacgagga gatcacagct cttaaaatta agtacaatcc
     3781 atttgcaaaa gctttccttg atgcaaagga aagaagtgat cacaaagaga tgatggagga
     3841 acccggagac agccagcaac ctgggtactc ccaatcaggg gggtggcttc ttcctggaac
     3901 cagcaccctg tgtccacctg caaatcctca tcctcagttt ggaggtgccc tctccctccc
     3961 ctccacgcac agctgtgaca ggtacccaac cctgaggagc caccggtcct caccctaccc
     4021 cagcccctat gctcatcgga acaattctcc aacctattct gacaactcac ctgcatgttt
     4081 atccatgctg caatcccatg acaattggtc cagccttgga atgcctgccc atcccagcat
     4141 gctccccgtg agccacaatg ccagcccacc taccagctcc agtcagtacc ccagcctgtg
     4201 gtctgtgagc aacggcgccg tcaccccggg ctcccaggca gcagccgtgt ccaacgggct
     4261 gggggcccag ttcttccggg gctcccccgc gcactacaca cccctcaccc atccggtctc
     4321 ggcgccctct tcctcgggat ccccactgta cgaaggggcg gccgcggcca cagacatcgt
     4381 ggacagccag tacgacgccg cagcccaagg ccgcctcata gcctcatgga cacctgtgtc
     4441 gccaccttcc atgtgagacc cagctttctt gtacaaagtg gttcgataac gaattccgcc
     4501 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     4561 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     4621 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     4681 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     4741 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     4801 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     4861 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     4921 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     4981 atgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     5041 ctttgaaaaa cacgatgata atatggccac aaccatgacc gagtacaagc ccacggtgcg
     5101 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     5161 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     5221 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5281 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5341 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5401 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5461 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5521 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5581 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5641 cacctggtgc atgacccgca agcccggtgc ctgatgcatc gatgtcgacc tgcagccaag
     5701 cttatcgatg gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5761 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5821 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5881 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5941 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6001 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6061 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6121 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6181 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6241 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6301 cccgcctgag atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6361 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     6421 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6481 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6541 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6601 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6661 aaatgaatat cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg
     6721 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6781 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7141 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     7201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7441 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7921 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7981 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8041 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8101 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8161 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8221 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8281 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8341 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8401 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8461 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8521 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8581 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8641 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8701 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     8761 gccgattcat taatgcagca agctcatggc tgactaattt tttttattta tgcagaggcc
     8821 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     8881 ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag cgggcagtga
     8941 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     9001 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     9061 ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     9121 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa
     9181 ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt
     9241 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     9301 ttaaatatgt actacaaact tagtagt
//
