LOCUS       pTMB303_ZF81187b        6638 bp DNA     circular SYN 02-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6638)
  AUTHORS   Tessa Bertozzi
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6638
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     118..188
                     /label=oTMB715_F
     primer_bind     123..182
                     /label=oTMB740
     primer_bind     123..182
                     /label=oTMB737
     primer_bind     123..182
                     /label=oTMB738
     primer_bind     123..182
                     /label=oTMB739
     misc_feature    144..166
                     /label=ZF81187bs_MM
     primer_bind     148..171
                     /label=oTMB836_F
     promoter        173..384
                     /label=pEFS
                     /label=EF-1-alpha promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=98.9"
                     /note="/match_length=99.5"
                     /note="/other=promoter"
     primer_bind     complement(366..405)
                     /label=oTMB715_R
     primer_bind     complement(373..384)
                     /label=Fragment 3.REV
     primer_bind     385..413
                     /label=oTMB716_F
     misc_feature    392..401
                     /label=Kozak
     misc_feature    401..490
                     /label=H3 tail ext
     misc_feature    491..610
                     /label=maxiflex40 (pENN175 codons)
     CDS             611..1249
                     /label=AsDnmt3L
                     /note="codon_start: 1 transl_table: 1 translation: 
                     MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVR
                     RDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNL
                     LLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQA
                     QVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL"
     primer_bind     611..639
                     /label=xENN318
     primer_bind     complement(611..638)
                     /label=xENN383
     primer_bind     complement(1038..1060)
                     /label=xENN347
     primer_bind     complement(1139..1158)
                     /label=oTMB736
     primer_bind     1222..1271
                     /label=xENN335
     primer_bind     complement(1222..1249)
                     /label=xENN336
     misc_feature    1250..1489
                     /label=XTEN80
     primer_bind     complement(1250..1272)
                     /label=xENN201
     primer_bind     1250..1269
                     /label=xENN202
     primer_bind     1489..1509
                     /label=oTMB724_F
     CDS             1490..1543
                     /codon_start=1
                     /label=bpNLS
                     /translation="KRTADGSEFESPKKKRKV"
     primer_bind     complement(1511..1569)
                     /label=xENN270
     primer_bind     complement(1517..1569)
                     /label=xENN277
     CDS             1544..2143
                     /codon_start=1
                     /label=ZFP 81187
                     /translation="GVPAAMAERPFQCRICMRNFSTSGNLTRHIRTHTGEKPFACDICG
                     RKFALSQDLNRHTKIHTHPRAPIPKPFQCRICMRNFSDRSALSRHIRTHTGEKPFACDI
                     CGRKFARNDDRKKHTKIHTGSQKPFQCRICMRNFSQSGNLARHIRTHTGEKPFACDICG
                     RKFAQRIDLTRHTKIHLRQKDAARGSGGDAKSLTAWS"
     primer_bind     complement(1567..1586)
                     /label=oTMB274_R
     primer_bind     2126..2163
                     /label=oTMB723_F
     misc_feature    2144..2194
                     /label=bpNLS
     primer_bind     complement(2170..2194)
                     /label=xENN222
     primer_bind     complement(2188..2208)
                     /label=oTMB716_R
     misc_feature    2195..2242
                     /label=XTEN16
     misc_feature    2243..2428
                     /label=KRAB
     primer_bind     complement(2329..2348)
                     /label=oTMB723_R
     misc_feature    2429..2437
                     /label=GCG linker
     CDS             2438..2464
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2462..2494)
                     /label=oTMB836_R
     misc_feature    2476..3084
                     /label=WPRE
     misc_feature    complement(2476..2500)
                     /label=JKN169
     misc_feature    complement(2539..2559)
                     /label=BP0156
     misc_feature    3104..3152
                     /label=synthetic polyA
                     /label=synthetic polyA from library vector
     primer_bind     3135..3181
                     /label=oTMB725_F
     misc_feature    3135..3154
                     /label=5' overlap
     primer_bind     complement(3277..3287)
                     /label=oTMB245
     primer_bind     complement(3871..3918)
                     /label=oTMB725_R
     repeat_region   3901..4030
                     /label=AAV2 ITR
     rep_origin      4116..4571
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
     promoter        4853..4957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4958..5818
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, 
                     andrelated antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             4958..5062
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAE"
     rep_origin      5989..6577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctact agtggatcag cagaccgatt gtgggctcta gagggcagag
      181 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      241 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      301 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      361 aacgggtttg ccgccagaac acagcctcag cgccaccatg gccaggacca aacaaactgc
      421 cagaaagagc accgggggga aagcgccccg aaaacagctg gcgaccaaag cagcccgcaa
      481 aagcgcaccg ggaggtgcaa gcagtggggc aggaagttcc tcaggaggaa gtgcggcggg
      541 atccggaagc ggtggcgcat caagtggcgc cgcaagtagc agtgccgggt ccgcagctgg
      601 ctccgggtcc tcccctgttg agatctacaa gaccgtgtcc gcctggaagc ggcagccaat
      661 cagagtgctg agcctgttcg gcaatattga caaggaactg aagtctctgg gcttcctgga
      721 aagcggcagc cacagcgagg gcggcacact gaagtacgtg gaagatgtga ccaacgtggt
      781 gcggagagat gtggaaaagt ggggaccttt cgacctgctg tacggcagca cccagcctct
      841 gggcggaagc tgcgacaggt gtcctggctg gtacatgttc caatttcata gaatcctgca
      901 gtacgccaga cctagacagg agtctcagcc cttcttctgg atcttcatgg ataacctgct
      961 gctcacagag gacgaccaga tgaccaccgt gcggttcctg caaacagagg ccgtcaccct
     1021 gcgcgacgtg agatctagag tgctgcagaa cgccgtccgg gtgtggagca acatccccgg
     1081 tcttaaaagc aagcacgagg ctctgtcccc aaaagaggaa gagagcctgc agggccacgt
     1141 gcggaccaga tctaagctgg ccgctcctaa agtggacccc ctggtgaaga actgcctgct
     1201 gcctctgaga gagtacttta agtatttcag ccagagcagc ctgcccctgg gaggacctag
     1261 cagcggcgcc ccacctccaa gcggaggctc tccagctggc agccctacca gcacagagga
     1321 aggcacaagc gagagcgcca cacctgagag cggccctggc acctccaccg agcccagcga
     1381 gggctctgcc cctggcagcc ccgccggaag ccccacctct acagaagaag gcaccagcac
     1441 agagccttct gaaggctctg cccctggaac atccaccgag cctagcgaga agcggactgc
     1501 cgatggatct gagtttgaga gccctaagaa aaaaagaaag gttggagtgc ctgctgctat
     1561 ggccgagaga ccatttcagt gcagaatctg catgcgcaat ttcagcacca gcggaaatct
     1621 cacaagacac atccggaccc ataccggcga gaagcccttt gcctgcgata tctgcggcag
     1681 aaagttcgct ctgtctcagg acctgaaccg gcacaccaag atccacacac accccagagc
     1741 ccctatcccc aagcctttcc agtgccggat ttgcatgcgg aacttcagcg acaggtccgc
     1801 cctgagcaga catatcagaa cacacaccgg cgaaaagcct ttcgcctgtg atatctgtgg
     1861 cagaaagttc gcccggaacg acgacagaaa aaagcacaca aagatccaca ccggatctca
     1921 gaaacctttc caatgtagaa tctgcatgag aaacttcagc cagagcggca acctggccag
     1981 acacatccgg acccacacag gcgaaaagcc tttcgcctgt gatatctgcg gccggaaatt
     2041 cgcccagaga atcgacctga cccggcacac caagatccac ctgagacaga aggacgccgc
     2101 cagaggcagc ggcggcgacg ccaagagcct gaccgcctgg tccaagagga ccgccgacgg
     2161 cagcgagttc gagcccaaga aaaagcggaa ggtgtccggc tctgaaaccc ctggcacaag
     2221 cgagtctgct acacctgagt ccagaaccct ggtcaccttc aaagacgtgt tcgtggactt
     2281 caccagagaa gaatggaagc tgctggatac cgctcagcag atcgtgtaca gaaacgtgat
     2341 gctggaaaac tacaagaatc tggtgtccct gggctaccag ctgaccaagc ctgacgtgat
     2401 cctgagactg gaaaagggcg aggaacccgg gagcggctat ccctatgacg tgcccgatta
     2461 tgcctgatag aattcgctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2521 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2581 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2641 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2701 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2761 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2821 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2881 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     2941 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3001 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3061 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctaataaaa tatctttatt
     3121 ttcattacat ctgtgtgttg gttttttgtg tggcggccgc tagtggcagc accagctcag
     3181 ggctggaggt ggtggctcct gagggtacct cagccccagg tggtggcccg ggaaccctgg
     3241 atgacagtgc caccatttgc cgtgtctgcc agaagccagg cgatctggtt atgtgcaacc
     3301 agtgtgagtt ttgtttccac ctggactgtc acctgccggc cctgcaggat gtaccagggg
     3361 aggagtggag ctgctcactc tgccatgtgc tccctgacct gaaggaggag gatggcagcc
     3421 tcagcctgga tggtgcagac agcactggcg tggtggccaa gctctcacca gccaaccagc
     3481 ggaaatgtga gcgtgtactg ctggccctat tctgtcacga accctgccgc cccctgcatc
     3541 agctggctac cgactccacc ttctccctgg accagcccgg tggcaccctg gatctgaccc
     3601 tgatccgtgc ccgcctccag gagaagttgt cacctcccta cagctcccca caggagtttg
     3661 cccaggatgt gggccgcatg ttcaagcaat tcaacaagtt aactgaggac aaggcagacg
     3721 tgcagtccat catcggcctg cagcgcttct tcgagacgcg catgaacgag gccttcggtg
     3781 acaccaagtt ctctgctgtg ctggtggagc ccccgccgat gagcctgcct ggtgctggcc
     3841 tgagttccca ggagctgtct ggtggccctg gtgatggccc ctgcccaact ttaagcttgc
     3901 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3961 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4021 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4081 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4141 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4201 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4261 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4321 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4381 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4441 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4501 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4561 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4621 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4681 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4741 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4801 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4861 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4921 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4981 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5041 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5101 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5161 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5221 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5281 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5341 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5401 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5461 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5521 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5581 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5641 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     5701 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5761 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5821 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6121 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6601 acggttcctg gccttttgct ggccttttgc tcacatgt
//
