LOCUS       Exported                7458 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 7458)
  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..7458
                     /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             1246..1302
                     /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             2092..2148
                     /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             2164..2187
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3010..3028)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3054..3278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3324..3752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3766..4096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3947..4082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4163..4957
                     /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    5131..5252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5301..5317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5325..5341
                     /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(5349..5379)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5394..5415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5703..6291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6462..7322)
                     /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(7323..7427)
                     /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 agctgagagt gttcctggcc
     1021 ctgcccatca tcatggtggt ggccttcagc atgtgcatca tctgcctgct gatggccggc
     1081 gacacctgga cagagacact ggcctacgtg ctgttctggg gcgtggccag catcggcacc
     1141 tttttcatca tcctgtacaa cggcaaggac ttcgtggacg cccccagact ggtgcagaaa
     1201 gagaagatcg accccgggct taaggagctc gcatgcggaa gcggagctac taacttcagc
     1261 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctagatctga attcatggtg
     1321 ggccggaaca gcgccatcgc cgcgggcgtg tgcggtgccc tcttcatagg gtactgcatc
     1381 tactttgacc gcaaaaggcg gagtgacccc aactccaaag gagaagaact gtttaccggc
     1441 gtggtgccaa ttctcgtgga actggatggc gatgtgaatg gccacaaatt ttctgtcaga
     1501 ggagagggtg aaggtgatgc cacaatcgga aagctcaccc tgaaattcat ctgcaccact
     1561 ggaaagctcc ctgtgccatg gccaacactg gtcactaccc tgacctacgg cgtgcagtgc
     1621 ttttccagat acccagacca tatgaagagg catgactttt tcaagagcgc catgcccgag
     1681 ggctatgtgc aggagagaac catctctttc aaagatgacg ggaaatacaa gacccgcgct
     1741 gtggtcaagt tcgaaggaga cacactggtg aatagaatcg agttgaaggg cacagacttt
     1801 aaggaagatg gaaacattct cggccacaag ctggaataca actttaactc ccacaatgtg
     1861 tacatcacag ccgacaagca aaagaatggc atcaaggcta acttcacagt cagacacaac
     1921 gtcgaggatg gaagcgtgca gctggccgac cattatcaac agaacactcc aatcggcgac
     1981 ggccctgtgc tcctcccaga caaccattac ctgtccaccc agacagtcct gagcaaagat
     2041 ccaaatgaaa aaggaacact cgagcttaag gagctcgcat gcggaagcgg agctactaac
     2101 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctag atctgaattc
     2161 atggactaca aggacgacga cgacaagtgc ggcctgttca tctgcccctg caacaagctg
     2221 aagtccagcg acgcctacaa gaaggcctgg ggcaacaacc aggatggcgt ggtggcttcc
     2281 cagcctgcca gagtgggaag cggcggaagc tctggcagca gaggctccga tggctctgac
     2341 ggcagcagca agggcgagga actgttcaca ggcgtggtgc ctatcctggt ggaactggac
     2401 ggcgacgtga acggccacaa gttttctgtg cggggagagg gcgaaggcga cgccacaatc
     2461 ggaaagctga ccctgaagtt catctgtacc accggcaagc tgcccgtgcc ctggcctaca
     2521 ctcgtgacca cactgggcta cggcctgcag tgcttcgcca gataccccga ccatatgaag
     2581 cggcacgact tcttcaagag cgccatgccc gagggctacg tgcaggatct gaccatcagc
     2641 ttcaaggacg atggcaagta caagaccaga gccgtcgtga agttcgaggg cgacaccctc
     2701 gtgaaccgga tcgagctgaa gggcaccgat ttcaaagagg acggcaacat cctgggccac
     2761 aagctggagt acaacttcaa cagccacaac gtgtacatca ccgccgacaa gcagaagaac
     2821 ggcatcaagg ccaacttcac cgtgcggcac aacgtggaag atggcagcgt gcagctggcc
     2881 gaccactacc agcagaacac ccccatcgga gatggccccg tgctgctgcc cgacaaccac
     2941 tacctgagct accagaccgt gctgagcaaa gatccaaatg aaaaaggaac ataatctaga
     3001 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     3061 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3121 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3181 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3241 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3301 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3361 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3421 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3481 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3541 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3601 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3661 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     3721 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3781 tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta
     3841 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3901 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3961 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4021 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4081 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4141 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4201 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4261 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4321 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4381 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4441 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4501 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4561 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4621 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4681 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4741 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4801 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4861 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4921 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4981 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5041 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5101 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5161 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5221 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5281 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5341 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5401 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5461 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5521 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5581 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5641 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5881 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5941 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6001 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6061 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6121 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6181 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6241 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6301 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6361 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6421 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6481 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6541 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6601 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6661 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6721 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6781 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6841 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6901 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6961 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7021 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7081 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7141 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7201 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7261 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7321 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7381 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7441 aaagtgccac ctgacgtc
//
