LOCUS       Exported                6900 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 6900)
  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..6900
                     /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             898..945
                     /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             1462..1518
                     /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"
     promoter        complement(2452..2470)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2496..2720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2766..3194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3208..3538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3389..3524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3605..4399
                     /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    4573..4694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4743..4759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4767..4783
                     /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(4791..4821)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4836..4857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5145..5733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5904..6764)
                     /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(6765..6869)
                     /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 gcttatgcgg
      901 gaccacatgg tgctgcacga gtacgtgaac gccgccggca tcaccggagg cgatggaggc
      961 tccggaggag gctctaagct gaacaattct ggcgcccctc acagggagaa gaggggagga
     1021 gagaccgacg agttctccaa tgtgcggaga ggccggggcc acagaatgca gcacctgtct
     1081 gagggcacaa agggaaggca agtgggctct ggaggcgacg gagagagatg gggcagcgat
     1141 aggggctctc gcggcagcct gaacgagcag atcgccctgg tgctgatgag gctgcaggag
     1201 gatatgcaga atgtgctgca gcgcctgcag aagctggaga ccctgacagc cctgcaggcc
     1261 aagagctcca ccagcacact gcagaccgca ccacagccta caagccagag gccatcctgg
     1321 tggccctttg agatgtcccc tggcgtgctg acattcgcca tcatctggcc cttcatcgcc
     1381 cagtggctgg tgtatctgta ctatcagagg cgccggagaa agctgaacgg atccctgaag
     1441 gagctggcat gcggcagcgg agcaaccaac ttctccctgc tgaagcaggc aggcgacgtg
     1501 gaggagaatc ccggccctcg gtctgaattc atgagcaagg gagaggagct gttcaccgga
     1561 gtggtgccta tcctggtgga gctggacggc gatgtgaatg gccacaagtt ttccgtgcgc
     1621 ggagagggag agggcgacgc aacaatcggc aagctgaccc tgaagttcat ctgcaccaca
     1681 ggcaagctgc ccgtgccttg gccaaccctg gtgaccacac tgggctacgg cctgcagtgt
     1741 tttgcccggt atccagacca catgaagaga cacgatttct ttaagtccgc catgcccgag
     1801 ggctacgtgc aggagaggac aatctctttc aaggacgatg gcaagtataa gacacgcgcc
     1861 gtggtgaagt ttgagggcga caccctggtg aaccggatcg agctgaaggg cacagacttc
     1921 aaggaggatg gcaatatcct gggccacaag ctggagtaca actttaattc ccacaacgtg
     1981 tatatcaccg ccgataagca gaagaacggc atcaaggcca atttcacagt gcggcacaat
     2041 gtggaggacg gctctgtgca gctggccgat cactaccagc agaacacccc tatcggcgac
     2101 ggccctgtgc tgctgcctga taatcactac ctgtcttatc agaccgtgct gagcaaggac
     2161 ccaaacgaga agggcacagg aggcgatgga ggcagcggag gaggctccaa gctgagagtg
     2221 ttcctggccc tgcccatcat catggtggtg gccttttcta tgtgcatcat ctgtctgctg
     2281 atggcaggcg acacctggac agagaccctg gcctacgtgc tgttttgggg cgtggccagc
     2341 atcggcacat tctttatcat cctgtataac ggcaaggact tcgtggatgc ccccaggctg
     2401 gtgcagaagg agaagatcga ttgactcgag catgcatcta gagggcccta ttctatagtg
     2461 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2521 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2581 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2641 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2701 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2761 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2821 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2881 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     2941 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3001 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3061 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3121 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3181 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3241 tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3301 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3361 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3421 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3481 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3541 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3601 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3661 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3721 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3781 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3841 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3901 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3961 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4021 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4081 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4141 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4201 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4261 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4321 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4381 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4441 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4501 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4561 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4621 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4681 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4741 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4801 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4861 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     5341 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5581 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5701 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5761 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5821 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5881 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5941 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6001 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6061 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6121 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6181 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6241 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6301 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6361 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6421 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6481 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6541 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6601 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6661 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6721 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6781 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6841 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
