LOCUS       Exported                7180 bp DNA     circular SYN 22-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7180)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Mar 24, 2022 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7180
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        7..507
                     /label=hPGK promoter
     primer_bind     510..559
                     /label=BamHI-Kozak-mKO2-F
     primer_bind     520..559
                     /label=Repair template 15
     CDS             530..1186
                     /codon_start=1
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     primer_bind     530..559
                     /label=T2A-mKO2-F
     primer_bind     1045..1075
                     /label=mKO2mut+
     primer_bind     complement(1045..1075)
                     /label=mKO2mut-
     primer_bind     complement(1152..1205)
                     /label=SalI-mKO2-BfuAI--R
     primer_bind     complement(1157..1187)
                     /label=Repair template 16
     primer_bind     complement(1158..1205)
                     
     primer_bind     complement(1201..1209)
                     /label=KS primer
     LTR             1292..1525
                     /label=3' LTR (Delta-U3)
     polyA_signal    1597..1718
                     /label=SV40 poly(A) signal
     primer_bind     complement(1597..1626)
                     /label=Repair template 16
     rep_origin      1758..1893
                     /label=SV40 ori
     promoter        complement(1914..1932)
                     /label=T7 promoter
     primer_bind     complement(1942..1958)
                     /label=M13 fwd
     rep_origin      2100..2555
                     /direction=RIGHT
                     /label=f1 ori
     promoter        2581..2685
                     /label=AmpR promoter
     CDS             2686..3546
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3717..4305
                     /direction=RIGHT
                     /label=ori
     protein_bind    4593..4614
                     /label=CAP binding site
     promoter        4629..4659
                     /label=lac promoter
     protein_bind    4667..4683
                     /label=lac operator
     primer_bind     4691..4707
                     /label=M13 rev
     promoter        4728..4746
                     /label=T3 promoter
     promoter        4774..5000
                     /label=RSV promoter
     LTR             5001..5181
                     /label=5' LTR (truncated)
     primer_bind     5190..5213
                     /label=HIV-2
     primer_bind     complement(5207..5227)
                     /label=HIV-1
     misc_feature    5228..5353
                     /label=HIV-1 Psi
     misc_feature    5846..6079
                     /label=RRE
     primer_bind     6416..6439
                     /label=XbaI-CTS-F
     primer_bind     6416..6439
                     /label=CTS
     primer_bind     6556..6605
                     /label=NheI-U6-F
     promoter        6606..6846
                     /label=U6 promoter
     primer_bind     6847..6905
                     /label=AgeI-Spscaffold-F
     primer_bind     6847..6901
                     /label=AgeI-BfuAI-F
     misc_RNA        6877..6962
                     /label=AU-sgRNA scaffold
     primer_bind     6877..6906
                     /label=P3-Block-F
     primer_bind     6877..6900
                     /label=P6-crCD45-Pr
     primer_bind     6877..6900
                     /label=P8-crCD45-Pr2
     primer_bind     6877..6898
                     /label=Bcl11b-5'short-F
     primer_bind     6877..6898
                     /label=Bcl11b-3'short-F
     primer_bind     6877..6898
                     /label=Bcl11b-Pr-P
     primer_bind     6877..6898
                     /label=Bcl11b-Pr-D
     primer_bind     complement(6952..7010)
                     /label=SFFV-sgRNA-R
     primer_bind     complement(6952..6976)
                     /label=XbaI-EcoRI-Sp_scaffold-R
     primer_bind     complement(6952..6975)
                     /label=EcoRI-Spscaffold-R
     primer_bind     complement(6952..6969)
                     /label=Opti-sgRNA scaffold R
     primer_bind     complement(6984..7010)
                     /label=NheI-Sp-R
     misc_feature    7011..7128
                     /label=cPPT/CTS
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccttaat gccgccacca tggtgagtgt
      541 gattaaacca gagatgaaga tgaggtacta catggacggc tccgtcaatg ggcatgagtt
      601 cacaattgaa ggtgaaggca caggcagacc ttacgaggga catcaagaga tgacactacg
      661 cgtcacaatg gccgagggcg ggccaatgcc tttcgcgttt gacttagtgt cacacgtgtt
      721 ctgttacggc cacagagtat ttactaaata tccagaagag ataccagact atttcaaaca
      781 agcatttcct gaaggcctgt catgggaaag gtcgttggag ttcgaagatg gtgggtccgc
      841 ttcagtcagt gcgcatataa gccttagagg aaacaccttc taccacaaat ccaaatttac
      901 tggggttaac tttcctgccg atggtcctat catgcaaaac caaagtgttg attgggagcc
      961 atcaaccgag aaaattactg ccagcgacgg agttctgaag ggtgatgtta cgatgtacct
     1021 aaaacttgaa ggaggcggca atcacaaatg ccaaatgaaa actacttaca aggcggcaaa
     1081 agagattctt gaaatgccag gagaccatta catcggccat cgcctcgtca ggaaaaccga
     1141 aggcaacatt actgagcagg tagaagatgc agtagctcat tcctaaagta gatgccgacg
     1201 tcgacctcga gggggggccc ggtaccttta agaccaatga cttacaaggc agctgtagat
     1261 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     1321 aaagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     1381 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1441 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1501 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     1561 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     1621 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     1741 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1801 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     1861 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     1921 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1981 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2041 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     2101 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2161 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2221 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2281 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2341 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2401 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2461 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2521 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     2581 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2641 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2701 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2761 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2821 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2881 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2941 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3001 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3061 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3121 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3181 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3241 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3301 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3361 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3421 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3481 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3541 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3601 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3661 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3721 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3781 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3841 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3901 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3961 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4021 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4081 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4141 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4201 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4261 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4321 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4381 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4441 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4501 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4561 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4621 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4681 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     4741 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     4801 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     4861 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     4921 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     4981 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     5041 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5101 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5161 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     5221 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     5281 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     5341 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     5401 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     5461 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     5521 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     5581 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     5641 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     5701 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     5761 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     5821 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     5881 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     5941 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     6001 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     6061 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     6121 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     6181 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     6241 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     6301 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     6361 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     6421 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     6481 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     6541 gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct
     6601 tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     6661 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     6721 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     6781 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     6841 aaggacgaaa caccggtgtc ttcgagacga agacccgttt aagagctatg ctggaaacag
     6901 catagcaagt ttaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     6961 gctttttttg aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     7021 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     7081 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     7141 agggacagca gagatccact ttggccgcgg ctcgaggggg
//
