LOCUS       Exported                7809 bp ds-DNA     circular SYN 12-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7809)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 30, 2020 from SnapGene Viewer 5.2.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7809
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             922..969
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1006..1272
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="MWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSF
                     NQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     CDS             1597..1653
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2443..2499
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2515..2538
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3361..3379)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3405..3629
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3675..4103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4117..4447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4298..4433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4514..5308
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5482..5603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5652..5668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5676..5692
                     /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)."
     promoter        complement(5700..5730)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5745..5766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6054..6642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6813..7673)
                     /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"
     promoter        complement(7674..7778)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccgccaccat gcgggaccac atggtgctgc acgagtacgt gaacgccgct
      961 ggcatcacag gcggagatgg cggatctggc ggcggaagca aactgatgtg gcacgaggga
     1021 ctggaagagg ccagcagact gtacttcggc gagcggaacg tgaagggcat gttcgaggtg
     1081 ctggaacccc tgcacgccat gatggaaaga ggcccccaga ccctgaaaga gacaagcttc
     1141 aaccaggcct acggccggga cctgatggaa gcccaggaat ggtgccggaa gtacatgaag
     1201 tccggcaatg tgaaggacct gacacaggcc tgggacctgt actaccacgt gttccggcgg
     1261 atcagcaagc agggctctga agccgccgct agagaagctg ctgctagagg cggagcttct
     1321 ggcgctggcg caggcgctgg ggccatcctg aatagcagag tgttcctggc cctgcccatc
     1381 atcatggtgg tggccttcag catgtgcatc atctgcctgc tgatggccgg cgacacctgg
     1441 acagagacac tggcctacgt gctgttctgg ggcgtggcca gcatcggcac ctttttcatc
     1501 atcctgtaca acggcaagga cttcgtggac gcccccaggc tggtgcagaa agagaagatc
     1561 gaccccgggc ttaaggagct cgcatgcgga agcggagcta ctaacttcag cctgctgaag
     1621 caggctggag acgtggagga gaaccctgga cctagatctg aattcatggt gggccggaac
     1681 agcgccatcg ccgcgggcgt gtgcggtgcc ctcttcatag ggtactgcat ctactttgac
     1741 cgcaaaaggc ggagtgaccc caactccaaa ggagaagaac tgtttaccgg cgtggtgcca
     1801 attctcgtgg aactggatgg cgatgtgaat ggccacaaat tttctgtcag aggagagggt
     1861 gaaggtgatg ccacaatcgg aaagctcacc ctgaaattca tctgcaccac tggaaagctc
     1921 cctgtgccat ggccaacact ggtcactacc ctgacctacg gcgtgcagtg cttttccaga
     1981 tacccagacc atatgaagag gcatgacttt ttcaagagcg ccatgcccga gggctatgtg
     2041 caggagagaa ccatctcttt caaagatgac gggaaataca agacccgcgc tgtggtcaag
     2101 ttcgaaggag acacactggt gaatagaatc gagttgaagg gcacagactt taaggaagat
     2161 ggaaacattc tcggccacaa gctggaatac aactttaact cccacaatgt gtacatcaca
     2221 gccgacaagc aaaagaatgg catcaaggct aacttcacag tcagacacaa cgtcgaggat
     2281 ggaagcgtgc agctggccga ccattatcaa cagaacactc caatcggcga cggccctgtg
     2341 ctcctcccag acaaccatta cctgtccacc cagacagtcc tgagcaaaga tccaaatgaa
     2401 aaaggaacac tcgagcttaa ggagctcgca tgcggaagcg gagctactaa cttcagcctg
     2461 ctgaagcagg ctggagacgt ggaggagaac cctggaccta gatctgaatt catggactac
     2521 aaggacgacg acgacaagtg cggcctgttc atctgcccct gcaacaagct gaagtccagc
     2581 gacgcctaca agaaggcctg gggcaacaac caggatggcg tggtggcttc ccagcctgcc
     2641 agagtgggaa gcggcggaag ctctggcagc agaggctccg atggctctga cggcagcagc
     2701 aagggcgagg aactgttcac aggcgtggtg cctatcctgg tggaactgga cggcgacgtg
     2761 aacggccaca agttttctgt gcggggagag ggcgaaggcg acgccacaat cggaaagctg
     2821 accctgaagt tcatctgtac caccggcaag ctgcccgtgc cctggcctac actcgtgacc
     2881 acactgggct acggcctgca gtgcttcgcc agataccccg accatatgaa gcggcacgac
     2941 ttcttcaaga gcgccatgcc cgagggctac gtgcaggatc tgaccatcag cttcaaggac
     3001 gatggcaagt acaagaccag agccgtcgtg aagttcgagg gcgacaccct cgtgaaccgg
     3061 atcgagctga agggcaccga tttcaaagag gacggcaaca tcctgggcca caagctggag
     3121 tacaacttca acagccacaa cgtgtacatc accgccgaca agcagaagaa cggcatcaag
     3181 gccaacttca ccgtgcggca caacgtggaa gatggcagcg tgcagctggc cgaccactac
     3241 cagcagaaca cccccatcgg agatggcccc gtgctgctgc ccgacaacca ctacctgagc
     3301 taccagaccg tgctgagcaa agatccaaat gaaaaaggaa cataatctag agggccctat
     3361 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3421 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3481 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3541 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3601 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3661 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4021 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     4081 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4141 tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4201 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4261 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4321 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4381 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4441 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     4501 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4561 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4621 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4681 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4741 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4801 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4861 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4921 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4981 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5041 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5101 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5161 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5221 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5281 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5341 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5401 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5461 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5521 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5581 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5641 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5701 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5761 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5821 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5881 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5941 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6061 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6121 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6241 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6481 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6721 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6781 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6841 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6901 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6961 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7021 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7081 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7141 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7201 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7261 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7321 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7381 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7441 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7501 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7561 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7621 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7741 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7801 cctgacgtc
//
