LOCUS       Crop-Seq-BFP-WPRE-TS-hU6-BsmBI       10353 bp    DNA     circular UNA 02-NOV-2021
DEFINITION  .
ACCESSION   urn.local...2z-attjxlb
VERSION     urn.local...2z-attjxlb
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    47..76
                     /note="Geneious type: motif"
                     /standard_name="switch oligo bind"
     promoter        87..327
     misc_RNA        2217..2292
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /standard_name="g RNA scaffold"
     LTR             2345..2525
     polyA_signal    2603..2724
     rep_origin      2764..2899
     promoter        complement(2920..2938)
     rep_origin      3106..3561
     promoter        3587..3691
     CDS             3692..4552
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="AmpR"
     rep_origin      4723..5311
     protein_bind    5599..5620
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
                     /standard_name="CAP binding site"
     promoter        5635..5665
     protein_bind    5673..5689
                     /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)."
                     /standard_name="lac operator"
     promoter        5734..5752
     promoter        5780..6006
     LTR             6007..6187
     misc_feature    6234..6359
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /standard_name="HIV-1 Psi"
     misc_feature    6852..7085
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to
                     thecytoplasm."
                     /standard_name="RRE"
     misc_feature    7495..7612
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /standard_name="cPPT/CTS"
     promoter        7751..8929
     intron          7982..8920
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /standard_name="EF-1-alpha intron A"
     gene            8942..9643
                     /standard_name="tagBFP"
     misc_feature    9659..10247
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /standard_name="WPRE"
     CDS             complement(10130..10141)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /standard_name="Factor Xa site"
ORIGIN      
        1 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccaagc agtggtatca
       61 acgcagagta catgggggga cccagagagg gcctatttcc catgattcct tcatatttgc
      121 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
      181 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
      241 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
      301 tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa tgagcacaca
      361 aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca aaatcttctt
      421 tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa acttttaata
      481 aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc ttcaccttta
      541 ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat cattttttga
      601 gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc tttaacaact
      661 gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc tgttgatgtt
      721 ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat ttcttctttt
      781 gttaataaac aattttccat aatacattta acaacatgtg atccagctgc tttttttaca
      841 gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc aatatttaat
      901 accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa tgcttttgta
      961 gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac atttgtgcct
     1021 tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt tgcaaaatca
     1081 aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc ttgttttaat
     1141 aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc atttttaatt
     1201 ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat tcaccaacac
     1261 ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca aatcatgact
     1321 tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt ccatcgatgt
     1381 agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt gctctcaatg
     1441 atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc aagtttaaag
     1501 tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct actaatgttt
     1561 tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct tgtattccat
     1621 ctttatctct agctgagtca tcaattacat atccataact ttcttcataa gcaaaaacaa
     1681 aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta aatccagtta
     1741 aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc aaatcacttg
     1801 ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt tttagatttg
     1861 ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt acaaaatgac
     1921 catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata ataataatat
     1981 ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca aattctggat
     2041 ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca acctcaataa
     2101 cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt ccattaactg
     2161 cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac gtctctgttt
     2221 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     2281 ccgagtcggt gcttttttaa gcttggcgta actagatctt gagacactgc tttttgcttg
     2341 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     2401 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     2461 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     2521 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     2581 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     2641 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2701 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     2761 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2821 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2881 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     2941 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     3001 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     3061 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     3121 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3181 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3241 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3301 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3361 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3421 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3481 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3541 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3601 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3661 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3721 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3781 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3841 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3901 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3961 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4021 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4081 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4141 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4201 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4261 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4321 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4381 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4441 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4501 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4561 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4621 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4681 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4741 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4801 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4861 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4921 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4981 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5041 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5101 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5161 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5221 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5281 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5341 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5401 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5461 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5521 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5581 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5641 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5701 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     5761 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     5821 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     5881 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     5941 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     6001 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6061 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6121 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6181 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     6241 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     6301 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     6361 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     6421 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     6481 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     6541 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     6601 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     6661 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     6721 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     6781 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     6841 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     6901 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     6961 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     7021 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     7081 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     7141 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     7201 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     7261 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     7321 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     7381 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     7441 tattcaccat tatcgtttca gaatcttgag acaaatggca gtattcatcc acaattttaa
     7501 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     7561 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     7621 ttacagggac agcagagatc cactttggcg ccggctcgag ggggcccggg tgcaaagatg
     7681 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
     7741 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     7801 gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc gcggggtaaa
     7861 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     7921 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     7981 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     8041 gccttgaatt acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     8101 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     8161 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     8221 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     8281 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     8341 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     8401 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     8461 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     8521 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     8581 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     8641 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     8701 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     8761 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     8821 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     8881 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgac gtacggccac
     8941 catgagcgag ctgatcaagg agaacatgca catgaagctg tacatggagg gcaccgtgga
     9001 caaccaccac ttcaagtgca ccagcgaggg cgagggcaag ccctacgagg gcacccagac
     9061 catgcggatc aaggtggtgg agggcggccc cctgcccttc gccttcgaca tcctggccac
     9121 cagcttcctg tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt
     9181 caagcagagc ttccccgagg gcttcacctg ggagcgggtg accacctacg aggacggcgg
     9241 cgtgctgacc gccacccagg acaccagcct ccaggacggc tgcctgatct acaacgtgaa
     9301 gatccggggc gtgaacttca ccagcaacgg ccccgtgatg cagaagaaaa ccctgggctg
     9361 ggaggccttc accgagacac tgtaccccgc cgacggcggc ctggagggcc ggaacgacat
     9421 ggccctgaag ctggtgggcg gcagccacct gatcgccaac atcaagacca cctaccggag
     9481 caagaagccc gccaagaacc tgaagatgcc cggcgtgtac tacgtggact accggctgga
     9541 gcggatcaag gaggccaaca acgagacata cgtggagcag cacgaggtgg ccgtggcccg
     9601 gtactgcgac ctgcccagca agctgggcca caagctgaac tgaacgcgtt aagtcgacaa
     9661 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     9721 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     9781 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     9841 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     9901 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     9961 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
    10021 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
    10081 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
    10141 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
    10201 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga
    10261 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
    10321 gaagggctaa ttcactccca acgaagacaa gat
//
