LOCUS       Exported                8815 bp ds-DNA     circular SYN 22-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8815)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Nov 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8815
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     misc_feature    772..792
                     /label=CMV -F
     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    2850..2870
                     /label=mU6-F
     misc_feature    2897..2918
                     /label=SV U6-F2
     misc_feature    2929..2931
                     /label=POSITION FOR INSERTING sHRNA (KD)
     misc_feature    2932..3006
                     /label=hCavin1 shRNA
     primer_bind     3008..3024
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3066..3099
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3670..3679
                     /label=Kozak seq
     misc_feature    3676..4851
                     /label=mCavin1 (isoform 1: NM_008986.2)
     misc_feature    3676..3678
                     /label=START
     misc_feature    4402..4476
                     /label=2 UC1
     misc_feature    4644..4672
                     /label=shRNA site - mutagenesis in purple
     regulatory      4864..4873
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4870..5589
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(4915..4936)
                     /label=EGFP-N
     CDS             5065..5077
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SHGV"
     misc_feature    complement(5176..5195)
                     /label=EXFP-R
     misc_feature    5523..5544
                     /label=EGFP-C
     protein_bind    5617..5650
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5706..6292
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6177..6188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6295..6311)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6821..7001
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7063..7651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7822..8682)
                     /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"
     misc_feature    8445..8464
                     /label=AMP-F
     promoter        complement(8683..8787)
                     /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 ttcaccttcc
     2941 acgtcaagaa gatccgcgag gttcaagaga cctcgcggat cttcttgacg tggaaggtgt
     3001 tttttctcga ggtcgacggt atcgataagc tcgcttcacg agattccagc aggtcgaggg
     3061 acctaataac ttcgtatagc atacattata cgaagttata ttaagggttc caagcttaag
     3121 cggccgctga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     3181 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     3241 gacagcagaa tatgggccaa caaggatatc tgtggtaagc agttcctgcc ccggctcagg
     3301 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     3361 gatgtttcca gggtgcccca aggacctgaa aatgaccctg tgccttattt gaactaacca
     3421 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     3481 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtat
     3541 tcccaataaa gcctcttgct gtttgcatcc gaatcgtgga ctcgctgatc cttgggaggg
     3601 tctcctcaga ttgattgact gcccacctcg ggggtctttc atcgaattct gcagtcgacg
     3661 gtaccgcggg ccaccatgga ggatgtcacg ctccatatcg ttgagcggcc gtattccgga
     3721 tttcccgatg cttcctcaga gggcccggag cccacccaag gggaggcgcg ggccacggag
     3781 gagccgtcgg ggaccggctc cgacgagctg atcaagtcgg accaggtgaa cggtgtgctg
     3841 gtgctgagcc ttctggataa aatcatcggc gccgttgacc agatccagct gacccaagcc
     3901 cagctggagg agcgacaggc ggagatggag ggcgctgtgc agagcatcca gggagagctg
     3961 agcaagctgg gcaaggcgca cgccaccacg agcaacaccg tgagcaagtt gctggagaag
     4021 gtgcgcaagg tcagcgtcaa cgtgaagacc gtgcgcggca gcctggagcg ccaggccggc
     4081 cagataaaga aactggaggt caacgaggcg gagctgctga ggcgccgcaa cttcaaagtc
     4141 atgatctacc aggatgaagt caagctgccg gccaaactga gcgtcagcaa gtcgctgaaa
     4201 gagtcggagg cactgcctga gaaggagggt gacgagctgg gcgagggcga gcggcccgag
     4261 gatgacaccg cggcgatcga gctgtcgtcc gacgaggcgg tggaggtgga ggaggtgatc
     4321 gaggagtccc gcgccgagcg catcaagcgc agcggcctgc ggcgcgtgga cgacttcaag
     4381 aaggccttct ccaaggagaa gatggagaag accaaggtgc gcacgcgtga gaacctggag
     4441 aagacgcgcc tgaagaccaa ggagaacctg gagaagacac ggcacacgct ggagaagcgc
     4501 atgaacaagc tgggcacgcg cctggtgccc gtggagcgac gagagaagct gaagacatcc
     4561 cgggacaagc tgcgcaagtc cttcacgccc gaccatgtgg tgtatgcgcg ctccaagacc
     4621 gctgtctaca aggtgccgcc tttcacattt catgttaaga aaattcgtga aggcgaggtg
     4681 gaggtgctga aggccaccga gatggtggag gtgggtcccg aggacgacga ggttggcgcg
     4741 gagcgcggcg aggccactga cctgctgcgc gggagcagcc ccgacgtgca cacgctgctg
     4801 gagatcaccg aggagtcgga cgccgtcctg gtggacaaga gcgacagcga cgatccaccg
     4861 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4921 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4981 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     5041 tggcccaccc tcgtgaccac cctgagccac ggcgtccagt gcttcagccg ctaccccgac
     5101 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5161 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5221 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5281 ctggggcaca agctggagta caacttcaac agccacaacg tctatatcat ggccgacaag
     5341 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5401 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5461 gacagccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     5521 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5581 tacaagtaac ctgcaggcaa ttcgtcgagg gacctaataa cttcgtatag catacattat
     5641 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
     5701 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5761 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5821 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5881 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5941 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     6001 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6061 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     6121 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6181 ccctcaatcc agcggacctt ccttcccggc ctgctgccgg ctctgcggcc tcttccgcgt
     6241 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     6301 cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     6361 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     6421 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6481 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6541 tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg
     6601 gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca
     6661 agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct
     6721 gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc
     6781 atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca
     6841 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6901 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6961 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca
     7021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7501 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
