LOCUS       Exported                9244 bp DNA     circular SYN 20-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9244)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Jun 7, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          5304..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2618..2931
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     misc_feature    2933..2985
                     /label=sh
     misc_feature    2933..2951
                     /label=hCortactin shRNA
                     /note="shRNA against human cortactin"
     misc_feature    2961..2979
                     /label=hCortactin shRNA
     primer_bind     2988..3004
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3046..3079
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        3108..3622
                     /label=5'LTR
     mRNA            3636..5285
                     /label=hCortactin 3D
     regulatory      5298..5307
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5304..6014
                     /label=mCherry
                     /note="mCherry"
     protein_bind    6042..6075
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    6131..6721
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6602..6613)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6724..6740)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             7250..7430
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7492..8080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8251..9111)
                     /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(9112..9216)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccgcg ctctagagat ccgacgccgc catctctagg
     2641 cccgcgccgg ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt
     2701 taatttgcat ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat
     2761 aatctgttct ttttaatact agctacattt tacatgatag gcttggattt ctataagaga
     2821 tacaaatact aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa
     2881 agtaattgtg tgttttgaga ctataaatat cccttggaga aaagccttgt ttgagaagca
     2941 cgagtcacag attcaagaga tctgtgactc gtgcttctct tttttctcga ggtcgacggt
     3001 atcgataagc tcgcttcacg agattccagc aggtcgaggg acctaataac ttcgtatagc
     3061 atacattata cgaagttata ttaagggttc caagcttaag cggccgctga aagaccccac
     3121 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa
     3181 ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa
     3241 caaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     3301 agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     3361 aggacctgaa aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     3421 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     3481 gccagtcctc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct
     3541 gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg tctcctcaga ttgattgact
     3601 gcccacctcg ggggtctttc atcgaattcg ccaccatgtg gaaagcttca gcaggccacg
     3661 ctgtgtccat cgcccaggat gacgcggggg ccgatgactg ggagactgac cctgattttg
     3721 tgaatgatgt gagtgagaag gagcaaagat ggggtgccaa gacggtgcag ggctccgggc
     3781 accaggagca tatcaacata cacaagctga gggagaatgt ctttcaagag catcagaccc
     3841 ttaaggagaa ggaacttgaa acaggaccaa aagcttccca tggctatgga gggaaatttg
     3901 gtgtggaaca agaccgaatg gataagtcag ctgtcggcca cgaatatcag tcgaaacttt
     3961 ccaagcactg ctcgcaggtg gactcggtcc gtggcttcgg aggcaagttt ggtgtccaga
     4021 tggacagagt tgatcagtct gctgtaggct ttgaatacca ggggaagact gagaagcatg
     4081 cctcccagaa agactactcc agtggttttg gcggcaagta tggcgtgcag gccgaccgag
     4141 tagacaagag cgcggtgggc ttcgactacc agggcaagac ggagaaacat gaatcccaga
     4201 gagattactc caaaggtttc ggcggcaaat acggtatcga caaggacaaa gtggataaga
     4261 gcgccgttgg ctttgagtat caaggcaaaa cggagaagca cgagtcccag aaagactatg
     4321 tgaaagggtt tggaggaaaa tttggtgtgc agacagacag acaagacaaa tgtgcccttg
     4381 gctgggatca ccaggagaaa ttgcagctgc atgaatccca aaaagattat aagactggtt
     4441 ttggaggcaa attcggtgtt cagtcggaga ggcaggactc cgctgctgtg gggtttgatt
     4501 acaaggagaa gctggccaag cacgagtccc agcaagacta ctccaaagga ttcggcggga
     4561 agtatggggt gcagaaggat cggatggata agaatgcgtc aacctttgag gatgtcaccc
     4621 aggtgtcctc tgcctaccag aagacagtac ctgtcgaagc tgtgaccagc aaaacaagta
     4681 acatcagagc taactttgaa aacctcgcta aggagaaaga gcaggaggac aggcggaagg
     4741 cggaggcgga gagagcccag cggatggcca aggagcggca ggagcaggaa gaggccagga
     4801 ggaagctgga ggagcaagcc agagccaaaa cgcaaacgcc ccctgtgtcg cccgcacctc
     4861 agccaaccga ggagaggctg ccctcgagcc ccgtcgatga ggatgcggct tccttcaagg
     4921 cagagctgag ctacagaggc cctgtgagtg ggacggagcc ggagcccgtg tacagcatgg
     4981 aggccgctga ctaccgagag gccagcagcc agcagggcct ggcctatgcc acagaggctg
     5041 tcgatgaaag cgcagaggcc ccgggccact atcccgcaga ggacagcacc gacgatgagt
     5101 acgagaacga tctggggatc acagccgtcg ccctgtacga ctaccaggct gcgggcgatg
     5161 atgagatctc atttgaccct gatgacatca tcaccaacat cgagatgatt gacgacggct
     5221 ggtggcgcgg ggtgtgcaag ggccggtacg ggctcttccc agccaactat gtggagctgc
     5281 ggcaggatcc accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca
     5341 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     5401 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     5461 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     5521 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     5581 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     5641 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     5701 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     5761 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     5821 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     5881 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     5941 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     6001 agctgtacaa gtaacctgca ggcaattcgt cgagggacct aataacttcg tatagcatac
     6061 attatacgaa gttatacatg tttaagggtt ccggttccac taggtacaat tcgatatcaa
     6121 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6181 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6241 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6301 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6361 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6421 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6481 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     6541 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6601 ttcggccctc aatccagcgg accttccttc ccgcgcggcc tgctgccggc tctgcggcct
     6661 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6721 catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca caagtagcaa
     6781 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     6841 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     6901 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     6961 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     7021 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     7081 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     7141 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     7201 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     7261 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7321 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7381 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc
     7441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9241 tgac
//
