LOCUS       Exported                5260 bp ds-DNA     circular SYN 29-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5260)
  AUTHORS   Tumor Cell Biology
  TITLE     Direct Submission
  JOURNAL   Exported Nov 26, 2019 from SnapGene 4.0.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5260
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             132..992
                     /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      1163..1751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2075..2105
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /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     2137..2153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2149..2205
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSELTLTKGNKSW"
     primer_bind     2174..2202
                     /label=Fragment.FOR
     promoter        2174..2192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2191..2228
                     /label=Fragment 1.FOR
     primer_bind     2203..2228
                     /label=Fragment 3.REV
     misc_feature    2205
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    2206..2228
                     /label=sgRNA
     misc_feature    2229..3029
                     /label=Untitled
     misc_feature    2229..2255
                     /label=hP16 C-KI 5' F
     misc_feature    2877..2896
                     /label=hp16 C GFP11 KI det F1
                     /note="hp16 C GFP11 KI det F1"
     misc_feature    3006..3029
                     /label=hp16 GFP11 C-terminal KI AsCpf1 guide
                     /note="hp16 GFP11 C-terminal KI AsCpf1 guide RNA-R1"
     primer_bind     complement(3009..3045)
                     /label=Fragment 1.REV
     misc_feature    3018..3029
                     /label=CDKN2A Exon3
                     /note="CDKN2A Exon3"
     misc_feature    3027..3029
                     /label=hp16 GFP11 C-terminal KI sgRNA-R1
                     /note="hp16 GFP11 C-terminal KI sgRNA-R1"
     misc_feature    3029
                     /label=hP16 3' F
     primer_bind     3031..3061
                     /label=Fragment 2.FOR
     primer_bind     3031..3056
                     /label=Fragment 2.FOR (2)
     misc_feature    3031..3038
                     /label=lenti-v6-CTCF-C-terminal KI
                     /note="lenti-v6-CTCF-C-terminal KI"
     misc_feature    3031..3038
                     /label=hCTCF exon 12
                     /note="hCTCF exon 12"
     misc_feature    complement(3031..3037)
                     /label=hCTCF-C-terminal-KI 5' HA cloning R
     misc_feature    3031..3036
                     /label=Pac1
                     /note="Pac1"
     misc_feature    3038..3103
                     /label=P2A sequence
                     /note="P2A sequence"
     misc_feature    3038
                     /label=P2A
                     /note="P2A"
     misc_feature    3104..3811
                     /label=mCherry coding sequence
                     /note="mCherry coding sequence"
     primer_bind     complement(3789..3812)
                     /label=Fragment 2.REV
     primer_bind     complement(3792..3819)
                     /label=Fragment 2.REV (2)
     primer_bind     3801..3829
                     /label=Fragment 1.FOR
     primer_bind     3805..3839
                     /label=Fragment 3.FOR
     misc_feature    3812..4290
                     /label=Untitled
     misc_feature    3812..4289
                     /label=CDKN2A Exon3
                     /note="CDKN2A Exon3"
     misc_feature    3812..3845
                     /label=hP16 3' F
     misc_feature    3812..3829
                     /label=hp16 GFP11 C-terminal KI sgRNA-R1
                     /note="hp16 GFP11 C-terminal KI sgRNA-R1"
     primer_bind     3812..3829
                     /label=Fragment 3.REV
     misc_feature    3812
                     /label=hp16 GFP11 C-terminal KI AsCpf1 guide
                     /note="hp16 GFP11 C-terminal KI AsCpf1 guide RNA-R1"
     misc_feature    4077..4097
                     /label=hp16 C GFP11 KI det R2
                     /note="hp16 C GFP11 KI det R2"
     misc_feature    4292..4584
                     /label=CDKN2A-AS1
                     /note="CDKN2A-AS1"
     misc_feature    complement(4519..4543)
                     /label=hP16 3' R3
     misc_feature    complement(4560..4584)
                     /label=hP16 3' R2
     primer_bind     complement(4585..4622)
                     /label=Fragment 3.REV
     primer_bind     complement(4585..4610)
                     /label=Fragment 1.FOR
     CDS             4608..5020
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="NSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSE
                     EARTDRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLA
                     TFAGFPRQALNRGLPLGFRFSALRHLDPKKLD"
     misc_feature    4608
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4617..4635)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4642..4658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4803..5258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagct cggaattaac cctcactaaa gggaacaaaa gctggccgat tgaaagaacc
     2221 agagaggcga gaagctaaag agtggtcgga gaatctaatg caagctacgg gagaaagaaa
     2281 ctcaagtgca aatgctgcct caggaataaa cgtaaaaaga gactttcaag tgcaaatgct
     2341 ccctcaggaa taaaataatc ttgagactct caagtgtaaa tgctgcctcg ggagaaccga
     2401 acggcgagct ggagcccata cgcaacgaga ttagagagga aggcagaagc cagagcacat
     2461 gaataaatga gcatccattt tgtttcagaa atgatcggaa accatttgtg ggtttgtaga
     2521 agcaggcatg cgtagggaag ctacgggatt ccgccgagga gcgccagagc ctgaggcgcc
     2581 ctttggttat cgcaagctgg ctggctcact ccgcaccagg tgcaaaagat gcctggggat
     2641 gcgggaaggg aaaggccaca tcttcacgcc ttcgcgcctg gcattgtgag caaccactga
     2701 gactcattat ataacactcg ttttcttctt gcaaccctgc gggccgcgcg gtcgcgcttt
     2761 ctctgccctc cgccgggtgg acctggagcg cttgagcggt cggcgcgcct ggagcagcca
     2821 ggcgggcagt ggactagctg ctggaccagg gaggtgtggg agagcggtgg cggcgggtac
     2881 atgcacgtga agccattgcg agaactttat ccataagtat ttcaatgccg gtagggacgg
     2941 caagagagga gggcgggatg tgccacacat ctttgacctc aggtttctaa cgcctgtttt
     3001 ctttctgccc tctgcagaca tccccgattt aattaacgga agcggagcta ctaacttcag
     3061 cctgctgaag caggctggag acgtggagga gaaccctgga cctatggtga gcaagggcga
     3121 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     3181 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     3241 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     3301 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     3361 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3421 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     3481 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3541 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3601 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3661 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3721 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3781 ctccaccggc ggcatggacg agctgtacaa gtgaaagaac cagagaggct ctgagaaacc
     3841 tcgggaaact tagatcatca gtcaccgaag gtcctacagg gccacaactg cccccgccac
     3901 aacccacccc gctttcgtag ttttcattta gaaaatagag cttttaaaaa tgtcctgcct
     3961 tttaacgtag atatatgcct tcccccacta ccgtaaatgt ccatttatat cattttttat
     4021 atattcttat aaaaatgtaa aaaagaaaaa caccgcttct gccttttcac tgtgttggag
     4081 ttttctggag tgagcactca cgccctaagc gcacattcat gtgggcattt cttgcgagcc
     4141 tcgcagcctc cggaagctgt cgacttcatg acaagcattt tgtgaactag ggaagctcag
     4201 gggggttact ggcttctctt gagtcacact gctagcaaat ggcagaacca aagctcaaat
     4261 aaaaataaaa taattttcat tcattcactc atttattgtc aacatttatt gagcacctat
     4321 tacaacaatt tcatcgcatg gaagacagca tcgtttctga cactgttgtt tcatgtatct
     4381 cttagaaaaa cgctgctatt agacatctaa cactatttat cttgaggtga taaaatatca
     4441 aaagccgtgt ctcaagatcg atgaaatgcg gttaaaatga tgaatagaaa ctctaggggg
     4501 acctcatatc gatagactcg agactggcac atctggagat ccgtatttat ccggcttccc
     4561 cttccagatc acgcgaggtt tggggcctct ctggttcttt caatcggcca attcgcccta
     4621 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4681 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4801 cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4861 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4921 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4981 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5041 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5101 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5161 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5221 taacgcgaat tttaacaaaa tattaacgtt tacaatttcc
//
