LOCUS       Exported                9964 bp ds-DNA     circular SYN 11-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9964)
  AUTHORS   Sascha
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 1, 2020 from SnapGene Viewer 5.1.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9964
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        286..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        2000..2240
                     /label=U6 Promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4130..4205
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the CRISPR/Cas9 system"
     misc_feature    4261..4378
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4517..5695
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4748..5686
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5695..5713)
                     /label=Vector.REV
     primer_bind     5699..5734
                     /label=Fragment.FOR (2)
     CDS             5714..6088
                     /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"
     primer_bind     complement(6071..6103)
                     /label=Fragment.REV (2)
     primer_bind     6089..6112
                     /label=Vector.FOR
     misc_feature    6104..6692
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6575..6586)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6764..6997
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7075..7196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      7236..7371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7392..7410)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7420..7436)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7578..8033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        8059..8163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8164..9024
                     /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"
     rep_origin      9195..9783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccagag agggcctatt tcccatgatt ccttcatatt tgcatatacg
     2041 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     2101 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     2161 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     2221 atatatcttg tggaaaggac gaaacaccgg agacggttgt aaatgagcac acaaaataca
     2281 catgctaaaa tattatattc tatgaccttt ataaaatcaa ccaaaatctt ctttttaata
     2341 actttagtat caataattag aatttttatg ttcctttttg caaactttta ataaaaatga
     2401 gcaaaataaa aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct ttaataatag
     2461 ctactccacc acttgttcct aagcggtcag ctcctgcttc aatcattttt tgagcatctt
     2521 caaatgttct aactccacca gctgctttaa ctaaagcatt gtctttaaca actgacttca
     2581 ttagtttaac atcttcaaat gttgcacctg attttgaaaa tcctgttgat gttttaacaa
     2641 attctaatcc agcttcaaca gctatttcac aagctttcat gatttcttct tttgttaata
     2701 aacaattttc cataatacat ttaacaacat gtgatccagc tgcttttttt acagctttca
     2761 tgtcttctaa aactaattca taatttttgt cttttaatgc accaatattt aataccatat
     2821 caatttctgt tgcaccatct ttaattgctt cagaaacttc gaatgctttt gtagctgttg
     2881 tgcatgcacc tagaggaaaa cctacaacat ttgttattcc tacatttgtg ccttttaata
     2941 attctttaca atagcttgtt caatatgaat taacacaaac tgttgcaaaa tcaaattcaa
     3001 ttgcttcatc acataattgt ttaatttcag ctttcgtagc atcttgtttt aataatgtgt
     3061 gatctatata tttgtttagt ttcatttttt ctcctatata ttcattttta attttaattc
     3121 tttaataatt tcgtctactt taactttagc gttttgaaca gattcaccaa cacctataaa
     3181 ataaattttt agtttaggtt cagttccact tgggcgaaca gcaaatcatg acttatcttc
     3241 taaataaaat tttagtaagt cttgtcctgg catattatac attccatcga tgtagtcttc
     3301 aacattaaca actttaagtc cagcaatttg agttaagggt gttgctctca atgatttcat
     3361 taatggttca atttttaatt tcttttcttc tggtttaaaa ttcaagttta aagtgaaagt
     3421 gtaatatgca cccatttctt taaataaatc ttctaaatag tctactaatg ttttattttg
     3481 ttttttataa aatcaagcag cctctgctat taatatagaa gcttgtattc catctttatc
     3541 tctagctgag tcatcaatta catatccata actttcttca taagcaaaaa caaaatttaa
     3601 tccgttatct tcttctttag caatttctct acccattcat ttaaatccag ttaaagtttt
     3661 tacaatatta actccatatt tttcatgagc gattctatca cccaaatcac ttgttacaaa
     3721 acttgaatat agagccggat tttttggaat gctatttaag cgttttagat ttgataattt
     3781 tcaatcaatt aaaattggtc ctgtttgatt tccatctaat cttacaaaat gaccatcatg
     3841 ttttattgcc attccaaatc tgtcagcatc tgggtcattc ataataataa tatctgcatc
     3901 atgtttaata ccatattcaa gcggtatttt tcatgcagga tcaaattctg gatttggatt
     3961 tacaacattt ttaaatgttt catcttcaaa tgcatgctct tcaacctcaa taacgttata
     4021 tcctgattca cgtaatattt ttggggtaaa tttagttcct gttccattaa ctgcgctaaa
     4081 aataattttt aaatcttttt tagcttcttg ctcttttttg tacgtctctg ttttagagct
     4141 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     4201 ggtgcttttt taagcttggc gtaactagat cttgagacaa atggcagtat tcatccacaa
     4261 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     4321 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     4381 ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggggg cccgggtgca
     4441 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
     4501 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     4561 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     4621 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     4681 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     4741 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     4801 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
     4861 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
     4921 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
     4981 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
     5041 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
     5101 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
     5161 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     5221 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     5281 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     5341 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     5401 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     5461 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     5521 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     5581 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     5641 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgtac
     5701 ggccaccttc gaaatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     5761 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     5821 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     5881 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     5941 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     6001 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     6061 gcacttcgtg gccgaggagc aggactgaac gcgttaagtc gacaatcaac ctctggatta
     6121 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6181 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6241 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6301 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6361 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6421 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6481 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     6541 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6601 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6661 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     6721 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     6781 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6841 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6901 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6961 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     7021 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     7081 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7141 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7201 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     7261 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     7321 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     7381 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     7441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     7501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     7561 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7621 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     7681 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     7741 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7801 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7861 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7921 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7981 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     8041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     8101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     8161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     8221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     8281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     8341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     8401 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     8461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     8521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     8581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     8641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     8701 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     8761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     9001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     9061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     9121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     9181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     9241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     9301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     9361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     9421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     9481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     9541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     9601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     9661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     9721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     9781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     9841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     9901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     9961 gaag
//
