LOCUS       Exported                7099 bp ds-DNA     circular SYN 30-MAY-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7099)
  AUTHORS   Venken Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 28, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7099
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(437..460)
                     /label=ASP15JAN01
     primer_bind     457..483
                     /label=ASP16AUG02
     primer_bind     460..483
                     /label=ASP14OCT47
     terminator      461..510
     CDS             complement(511..1521)
                     /codon_start=1
                     /gene="SpnR"
                     /label=SpmR
                     /translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPRASKQQAR
                     YAVGRCLMLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHLEPTLLAV
                     HLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTI
                     VVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPA
                     AEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVA
                     ADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1522..1655)
                     /label=Spm promoter
     primer_bind     complement(1637..1659)
                     /label=ASP14OCT46
     primer_bind     complement(1638..1659)
                     /label=ASP16AUG01
     primer_bind     1656..1681
                     /label=ASP14NOV51
     rep_origin      1694..2376
                     /direction=RIGHT
                     /label=ColE1 origin
     primer_bind     complement(2438..2461)
                     /label=ASP14NOV50
     misc_feature    2466..2495
                     /label=STUFFER
     primer_bind     2504..2527
                     /label=ASP14NOV104
     misc_feature    2534..2573
                     /label=Barcode 9
     primer_bind     complement(2580..2607)
                     /label=ASP14NOV103
     primer_bind     2604..2626
                     /label=ASP14NOV60
     primer_bind     2607..2626
                     /label=ASP15APR19
     polyA_signal    2608..2803
                     /label=SV40 Late
     primer_bind     2667..2693
                     /label=ASP14NOV62
     polyA_signal    2669..2797
                     /label=SV40 Early (rev)
     primer_bind     complement(2774..2797)
                     /label=ASPNOV61
     primer_bind     complement(2779..2807)
                     /label=ASP14NOV59
     CDS             complement(2808..4871)
                     /codon_start=1
                     /label=White+
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLLPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     primer_bind     4866..4886
                     /label=NM16NOV20
     promoter        complement(4875..5029)
                     /label=DSCP
     misc_feature    4938..4943
                     /label=DPE
                     /note="Downstream Promoter Element"
     misc_feature    4944..4953
                     /label=MTE
                     /note="Motif Ten Element"
     misc_feature    4967..4972
                     /label=Inr
                     /note="Inititator"
     TATA_signal     4996..5001
                     /label=TATA
     primer_bind     complement(5014..5029)
                     /label=NM17JAN03
     primer_bind     complement(5014..5029)
                     /label=NM16NOV19
     primer_bind     5030..5054
                     /label=ASP15APR06
     enhancer        5068..5102
                     /label=GMR Enhancer
     enhancer        5103..5137
                     /label=GMR Enhancer
     enhancer        5138..5172
                     /label=GMR Enhancer
     enhancer        5173..5207
                     /label=GMR Enhancer
     enhancer        5208..5242
                     /label=GMR Enhancer
     primer_bind     complement(5243..5270)
                     /label=ASP15APR05
     polyA_site      5276..5323
                     /label=HSV-TK PolyA
                     /note="No BCD"
     primer_bind     5324..5351
                     /label=Mock NM15SEP15
     CDS             complement(5328..5927)
                     /codon_start=1
                     /product="Puromycin Resistance Gene pac. Encodes for 
                     Puromycin N-Acetyltransferase and confers resistance to 
                     puromycin in eukaryotic and prokaryotic cells. Is NOT good 
                     at selecting Gram Negative bacteria like E.Coli. 125ug/ml 
                     and precise pH adjustments required. "
                     /label=pac
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPVWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5908..5928)
                     /label=Mock NM15SEP14
     primer_bind     5925..5946
                     /label=NM15OCT02
     misc_feature    5929..5931
                     /label=Kozak
     mRNA            complement(5936..5941)
                     /label=SD
     promoter        complement(5942..6009)
                     /label=CP6 Promoter
     primer_bind     complement(5989..6013)
                     /label=ASP14SEP09
     primer_bind     6007..6039
                     /label=NM16NOV21
     promoter        complement(6014..6468)
                     /label=Hsp70B
     primer_bind     6014..6039
                     /label=ASP14NOV42
     primer_bind     complement(6338..6359)
                     /label=ASP14NOV41
     primer_bind     6355..6380
                     /label=ASP14NOV40
     primer_bind     complement(6447..6475)
                     /label=ASP14NOV39
     primer_bind     complement(6459..6475)
                     /label=NM16NOV22
     primer_bind     6477..6505
                     /label=ASP16SEP18
     primer_bind     6477..6505
                     /label=ASP16SEP16
     primer_bind     6480..6500
                     /label=ASP14NOV46
     protein_bind    complement(6481..6592)
                     /label=attB_TT
                     /bound_moiety="phage phi-C31 integrase"
     misc_feature    complement(6514..6547)
                     /label=attB_TT
     primer_bind     complement(6556..6592)
                     /label=ASP14JUL13
     primer_bind     complement(6571..6596)
                     /label=ASP16SEP17
     primer_bind     complement(6571..6596)
                     /label=ASP16SEP15
     primer_bind     complement(6573..6592)
                     /label=ASP14NOV45
     primer_bind     6599..6632
                     /label=ASP15MAR25
     CDS             complement(6710..6997)
                     /codon_start=1
                     /label=LacZ
                     /translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGE"
     primer_bind     6850..6866
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6873..6891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6894..6950
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(6957..6975)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        complement(7029..7076)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(7071..7087)
                     /label=ASP15MAR27
     primer_bind     complement(7073..7099)
                     /label=ASP15MAR13
     primer_bind     join(7095..7099,1..32)
                     /label=ASP15JUL28
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgctggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaca tgaagtttta aatcaatcta
      481 aagtatatat gtgtaacatt ggtctagtga ttatttgccg actaccttgg tgatctcgcc
      541 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaaac gatcttcttc
      601 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
      661 ctccattgcc cagtcggcag ctacatcctt cggcgcgatt ttgccggtta ctgcgctgta
      721 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
      781 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
      841 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
      901 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
      961 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     1021 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     1081 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     1141 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     1201 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     1261 tgatagttga gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt
     1321 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     1381 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc
     1441 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
     1501 ggaccagttg cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta
     1561 tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg
     1621 tagcagcgaa gtcgaggcat ttctgtcctg gctggtcaca aaaggatcta ggtgaagatc
     1681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtgtc
     2461 aagcgcagct gcccgggtct agagtcgacg catgcctcct gacgttctcc ttgttctcca
     2521 cggtcttaag cttagcacga gcgatcgaga gagatagaga gcgagataga gcgaagcttg
     2581 ttaattcgtc gagcgcttgc catactgaaa aaacctccca cacctccccc tgaacctgaa
     2641 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2701 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2761 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaagctca ctccttgcga
     2821 cgagcgcgca ggcgcagagc cagataggcc agcacgcgga aggacacaat caggatggcc
     2881 aggcccacat aatccagtgg caggtcggcg gcactgaaat tcagtgtctc caggatgacc
     2941 tttccgctgc tggggcaggt cgtgttgctg ctggtgcagg agatctcgcc gggctccaca
     3001 tcggcccact gattgatcag caggccctca ttggcgtagc gaaaccagct caggtagctc
     3061 agccacttca ggtacacggg cacgctgccg gagttcagaa agaatccgcc gaacagcagg
     3121 aaggggataa tcacgggggg tcccacgctc agggccatgc tggtgctgct ggaggcgcag
     3181 ctgatcaggt agccgaagct ggtcgacacg ttggccacca gggtcaccag ggccaggcaa
     3241 ttaaagaaat gcagcactcc ggcacgcagg ccaatcatgg ggtaggcaat ggcggtaaac
     3301 accaggggca cggtcaggaa cagcggcagc tcggcaatgg tctttcccag gaagtaggta
     3361 tcgcagcggt acaggcggct gcgggcctcg cgcatgaaca cgggcagctc gctggtgaac
     3421 acgttgatgg tggcgaacac gttctggaac gtcatgttcg tcagaaacag aaaaatggct
     3481 ccattgatgt tcatcactcc gacctgcgtc agctgctgtc ccaggaagat caggccgatc
     3541 agaatggcca ccatcgtggt ctggatcagg cggaccttga ccagcagggg ctccttcagc
     3601 acactcagcc agctgcgcca cagcacggcg cgaaactgca taaaccaggt agccttatac
     3661 gtatagccgt tctcgggctg ctccaggggc ttctccaggt tcttcgtggc cagcagctgc
     3721 tccatgtcgc gggccacctt ggagatggcg aagttatcgc agatcttggc aatgcgatcg
     3781 cggctctcaa tctcgcgtcc tggcaccacg gccagcactt gcacatagaa gtcggcgggg
     3841 ttgtaattcg tggggcactg ggcgcccaca tagctgaaga aatccacggc ctcgcttggg
     3901 gttcccagga aggccacgcg tccctcggcc atcagcagaa tcttatcaaa cagctcgaac
     3961 agctccgagc tgggctggtg gatcgtcaga atgaccgtct ttcccttctg gctcagcttc
     4021 ttcagcacct gcaccacgga gtgggcggtg aagctatcca ggccgctggt cggctcgtcg
     4081 caaatcagca gggggggatc ggtcagggcc tcgctggcaa aggccaggcg cttgcgctct
     4141 ccgccactca ggcccttcac gcggcctggc acgccaataa tggtgtgctg gcacttgctc
     4201 aggctcagct cctggatgac ctggtccacg cgggccacgc gctgacggta tgtcaggtgg
     4261 cggggcatgc ggaccatggc ctgaaagatc agatgctcgc gggctgtcag gctgccgatg
     4321 aacagatcgt cctgctgcac gtaggcgcag cgggcctgca tctccttggc atccacgggc
     4381 tggccgttca gcaggcgcat gccgcttggg gacacctgaa tgccctgggg gctgcggaag
     4441 gccagggcgt tcagcagcgt ggtctttccg gcgccactac ttcccatcac ggccagcagc
     4501 tcgccggggt aggccactcc gcacacattc ttcagcaggt gcttgcgtgg ggctgggata
     4561 tggcgctcat tgcagaacag tccgcgggtg cggttcacca gctgacgcca gccactgcct
     4621 ggctggttca cggcgccgaa gatatccatg ttatgccagg cgtacgtcag attctcggcc
     4681 agctgtccgg atccgcttcc gctatcctct ggggggctgg ggggcagcag ggttccgtaa
     4741 ttcttggcct ggccaaatcc ctgattgatg cacgactggc tggcggctcc gctatcgccg
     4801 ttgttcaggt gctcggcgct tgggtgcttg gagcctccgc ggatcagcag ctcctggtcc
     4861 tcctgtccca tttggtttgg tatgcgtctt gtgattcaaa gttggcttat tcaaaggata
     4921 ttaacgaagg cagcggcacg tctgctcggc tcgaacattc attcacaact gatgctctca
     4981 gccaccccgc gcccttttat accgctgcgc tcgatccccg ggcgagctcg ggaggggcat
     5041 gacgtaccta cctcgactgc tactcgactt aaaggcattt caagggtttc cactgggctc
     5101 gacttaaagg catttcaagg gtttccactg ggctcgactt aaaggcattt caagggtttc
     5161 cactgggctc gacttaaagg catttcaagg gtttccactg ggctcgactt aaaggcattt
     5221 caagggtttc cactgggctc gacagtcagg tgctagtcgt acgctcctga cagcggaaca
     5281 aacgacccaa cacccgtgcg ttttattctg tctttttatt gccaagctta ggctcctggc
     5341 ttacgggtca tacaccaggt gcgtgggccc tcgggcacct ccacatcggc cgtcacggtg
     5401 aagcccagac gctcgtagaa gggcagattg cgtggggcac tggtttccag gaaggctggc
     5461 actccagcac gctcggcagc ttccactcct ggcagcacca cggcactgcc caggcccttg
     5521 ccctggtggt cggggctcac gcccacggtg gccaggaacc agacgggctc ctttggacgg
     5581 tgtggggcca gcaggccctc catctgctgc tgggcagcca gacgggagcc actcagctcg
     5641 gccatacggg gtccaatctc ggcgaacacg gctccagcct ccacgctctc gggggtggtc
     5701 cacacggcca cggcagctcc atcatcggcc acccagacct tgccgatatc caggcccacg
     5761 cgtgtcagga acagctcctg gagctcggtc acacgctcaa tgtgacggtc tgggtccacg
     5821 gtgtgacggg tggctgggta atcggcgaag gcagcagcca gggttcgcac ggcacggggc
     5881 acatcatcac gggtggccag acgcaccgtg ggcttatact cggtcatttt gttttcctcc
     5941 ttatgttaag cttactcagt tattatatca taaatatctg tgtcaagaat aaactcccac
     6001 atggattcga gtattgagag ttctcttctt gtattcaata attacttctt ggcagatttc
     6061 agtagttgca gttgatttac ttggttgctg gttactttta attgattcac tttaacttgc
     6121 actttactgc agattgttta gcttgttcag ctgcgcttgt ttatttgctt agctttcgct
     6181 tagcgacgtg ttcactttgc ttgtttgaat tgaattgtcg ctccgtagac gaagcgcctc
     6241 tatttatact ccggcgctct tttcgcgaac attcgaggcg cgctctctcg aaccaacgag
     6301 agcagtatgc cgtttactgt gtgacagagt gagagagcat tagtgcagag agggagcgac
     6361 ccaaaaagaa aagagagaat aacgaataac ggccagagaa atttctcgag ttttctttct
     6421 gccaaacaaa tgacctacca caataaccag tttgttttgg gattctagct cctgacagcg
     6481 catcatgatg gaccagatgg gtgaggtgga gtacgcgccc ggggagccca agggcacgcc
     6541 ctggcacccg caccgcggct tcgagacagt gacctacatc gtcgaccaat tcctccctgg
     6601 gatccggaga gagaccctat gcggcatcag agcagattgt actgagagtg caccatatgc
     6661 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgt cattcgccat
     6721 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6781 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     6841 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattcg
     6901 agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt gagtattcta
     6961 tagtgtcacc taaatagctt ggcgtaatca tggtcatgat ctgcctcctg ggtttagttc
     7021 ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt gtcaacaggg
     7081 gtctctcgct gatccgtta
//
