LOCUS       Exported                4774 bp ds-DNA     circular SYN 07-DEC-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4774)
  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..4774
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             complement(174..1034)
                     /codon_start=1
                     /label=AmpR
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1035..1139)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1217..1672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1658..1679)
                     /label=NM15SEP05
     primer_bind     complement(1660..1683)
                     /label=NM15OCT37
     primer_bind     1677..1708
                     /label=NM16FEB01
     primer_bind     1677..1705
                     /label=NM15DEC05
     primer_bind     1680..1709
                     /label=NM15OCT31
     primer_bind     1684..1705
                     /label= M16OCT08
     terminator      complement(join(1698..1725,1751..1775))
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1762..1778)
                     /label=NM15DEC06
     primer_bind     1775..1799
                     /label=NM15DEC07
     misc_feature    1776..1865
                     /label=spy Term
     primer_bind     complement(1841..1865)
                     /label=NM16FEB04
     primer_bind     complement(1842..1866)
                     /label=NM16AUG01
     primer_bind     complement(1842..1865)
                     /label=NM15DEC08
     primer_bind     1867..1883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1884..1910)
                     /label=NM16OCT09
     promoter        1890..1908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1947..1971
                     /label=NM20NOV01
     CDS             1953..3770
                     /codon_start=1
                     /label=FC31_HS
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     primer_bind     1997..2018
                     /label=FC31-SEQ-1
     primer_bind     complement(2467..2490)
                     /label=FC31-SEQ3
     primer_bind     2610..2633
                     /label=FC31_Seq
     primer_bind     complement(2960..2981)
                     /label=FC31-SEQ2
     primer_bind     complement(3755..3779)
                     /label=NM20NOV02
     primer_bind     3774..3801
                     /label=NM15MAY02
     primer_bind     3774..3801
                     /label=NM15MAY01
     primer_bind     3811..3839
                     /label=NM16OCT10
     promoter        complement(3811..3829)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3845..3861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3861..3882
                     /label=NM16AUG02
     primer_bind     3863..3885
                     /label=NM15DEC09
     primer_bind     3863..3885
                     /label=NM16FEB03
     misc_feature    complement(3864..3924)
                     /label=L3S2P21 Term
                     /note="Synthetic Terminator. Very Strong. See Chen, 2013 
                     (Chris Voigt)"
     primer_bind     complement(3899..3924)
                     /label=NM15DEC10
     primer_bind     3925..3949
                     /label=NM15DEC11
     misc_feature    3934..3978
                     /label= rrnB T1 Terminator
     primer_bind     complement(3978..4015)
                     /label=NM16FEB02
     primer_bind     complement(3978..4003)
                     /label=NM15DEC12
     primer_bind     complement(3988..4003)
                     /label=NM16OCT11
     primer_bind     complement(3991..4007)
                     /label=NM15OCT32
     primer_bind     4001..4030
                     /label=NM15OCT38
     primer_bind     4001..4030
                     /label=NM15SEP06
     rep_origin      complement(join(4189..4774,1..3))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
       61 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
      121 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
      181 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
      241 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
      301 gcaatgatac cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      361 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
      421 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
      481 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      541 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      601 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      661 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      721 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      781 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      841 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      901 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      961 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1021 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1081 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     1141 acatttcccc gaaaagtgcc acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1201 aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1261 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     1321 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     1381 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcgc tggcccacta
     1441 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     1501 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     1561 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     1621 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgt
     1681 aacattgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc
     1741 tggcagttta tggcgggcgt cctgcccgcc accctttcag ccaaaaaact taagaccgcc
     1801 ggtcttgtcc actaccttgc agtaatgcgg tggacaggat cggcggtttt cttttctctt
     1861 ctcaatgtaa aacgacggcc agtgaattgt aatacgactc actatagggt gaattccgct
     1921 taggacctta gatattgatc tcaaggtctc gaatggacac ctatgccggt gcctacgaca
     1981 gacagtcccg ggagcgggaa aactccagtg cagccagccc cgccacgcaa cggtcagcta
     2041 acgaggataa ggcggccgac ctgcagaggg aggtggagag ggacggcggc agattcagat
     2101 tcgtaggcca ctttagcgag gcaccaggga ctagtgcctt tgggacagca gagaggccag
     2161 agttcgagag aatcctgaat gagtgccgag cgggccgcct gaacatgatc atagtttacg
     2221 atgtttcccg gttctcacgg ctgaaggtta tggacgccat ccctatcgta agtgagctgc
     2281 tggcactcgg ggtgactatt gttagcactc aggagggagt gttccggcag ggaaatgtca
     2341 tggacctgat tcatttgatt atgcgcctgg acgccagcca caaagaaagc agccttaaga
     2401 gcgcgaaaat cttggacaca aagaatcttc agcgagagct tggcggctac gtgggaggga
     2461 aggcccccta cgggttcgaa ctcgttagcg agacaaaaga aatcaccaga aacggccgaa
     2521 tggtcaacgt cgtcattaat aaactggcac attccacaac cccactgact gggccatttg
     2581 aattcgaacc cgacgtgatc cgttggtggt ggcgtgaaat taagacgcac aagcatctgc
     2641 cctttaaacc tggatctcag gcagccattc acccgggctc cattactggg ctttgtaagc
     2701 ggatggacgc cgacgcagtc ccaacgaggg gggagacaat aggcaaaaag actgcctcaa
     2761 gcgcgtggga ccctgccaca gtcatgcgca ttctccggga tccccgcatt gccgggttcg
     2821 ccgcggaggt gatatataaa aagaagcccg acggtacacc cactacaaag attgagggct
     2881 accggatcca gcgcgacccc attaccctgc ggccagtcga actggattgc gggcctatta
     2941 tcgaaccggc agaatggtac gaacttcagg cctggctgga tggacggggg agaggcaaag
     3001 ggctatctag aggccaggct atcttgtccg cgatggacaa actttattgc gaatgcggcg
     3061 ccgtcatgac atccaagaga ggcgaagaga gcatcaaaga tagttataga tgcagacgtc
     3121 gcaaagtggt cgatccctca gctcccggac agcacgaggg cacctgtaat gtcagtatgg
     3181 ccgcattaga caagttcgtc gcagaacgta tctttaacaa gatccgtcac gctgaaggag
     3241 atgaagaaac cctggctctg ttatgggagg ccgccagacg atttgggaag ctgacagagg
     3301 ctccagaaaa atctggcgaa cgtgccaacc tggtagctga gagagcggat gcgctcaatg
     3361 cactggagga actctacgag gatagggcag cgggcgccta cgatgggcca gtgggtcgca
     3421 agcacttccg gaaacagcag gcagcactga cacttagaca acagggcgcc gaggagaggc
     3481 tcgccgagct agaggccgct gaggccccga agctccccct ggatcagtgg ttccctgagg
     3541 atgccgacgc tgatcccacc ggacctaaaa gctggtgggg gcgagcttcc gtggacgata
     3601 aacgggtttt cgtcggcctc ttcgtggaca agatcgtggt gactaagagc accactggca
     3661 gagggcaagg cacaccaatc gagaagcgcg catctatcac ttgggctaag cctcctaccg
     3721 acgacgatga ggatgacgca caggacggga cagaggatgt agcagcctaa gcttcgagac
     3781 cactcatcgc ccaatcacta gtgaattcgt tctatagtgt cacctaaata ccatagagcc
     3841 gttggtcata gctgtttcct gtcggaccaa aacgaaaaaa ggcccccctt tcgggaggcc
     3901 tcttttctgg aatttggtac cgagcaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3961 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctgacctt ccgcttcctc
     4021 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4081 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4141 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4201 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4261 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4321 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4381 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4441 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4501 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4561 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4621 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4681 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4741 caagcagcag attacgcgca gaaaaaaagg atct
//
