LOCUS       Exported                7717 bp ds-DNA     circular SYN 12-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7717)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Apr 12, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7717
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        175..554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        555..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate earlypromoter"
     promoter        803..821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             844..867
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an 
                     enterokinasecleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             871..894
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an 
                     enterokinasecleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             898..921
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an 
                     enterokinasecleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    943..3213
                     /label=mLIG3-179-936
     polyA_signal    3257..3481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3527..3955
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
     promoter        3969..4298
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4149..4284
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4365..5159
                     /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    5333..5454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5527..5543
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5551..5581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5596..5617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
     rep_origin      complement(5905..6490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6661..7521)
                     /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(7522..7626)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
       61 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      121 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      181 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      541 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      721 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      781 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      841 atggattaca aggacgatga cgataagggt gattacaagg acgatgacga taagggtgat
      901 tacaaggacg atgacgataa gcttggtacc gagctcggat ccagccagca tattgcagac
      961 ctgtcttcta aggcagctgc cacacctaag aagaaagctg ctgtccaggc taagctgaca
     1021 accactggcc aggtgacatc tccagtgaaa ggtgcttcgt ttattaccag taccaatcct
     1081 cggaagttct ctggcttttc agcagcaaaa cccaacaact ctgagcaagc cccctcaagc
     1141 cctgctcctg gaacgagtct gtctgcaagt aaatgtgacc ctaagcacaa agattgtctg
     1201 ctacgagagt tccggaagct gtgcgccatg gtggctgaaa atcctagcta taacaccaag
     1261 acccagatca tccatgactt cttgcagaaa ggctccacag gagatggctt tcacggtgat
     1321 gtgtacctga cagttaagct gctgctgcct ggtgtcatca agagtgttta caacttgaat
     1381 gataagcaga ttgtaaaact ttttagccga atttttaact gcaacccaga tgatatggcc
     1441 cgggacctag aacagggtga cgtgtcagag acgatcagaa tcttctttga gcagagcaag
     1501 tctttccccc cagctgccaa aagcctcctc accattcagg aagtggatgc cttcctcctg
     1561 cacctctcca agctcaccaa agaggatgag cagcagcagg ccctgcagga catagcctcc
     1621 cggtgtacag ccaatgacct taagtgcatt attcggctga tcaaacatga tctgaagatg
     1681 aactcgggtg caaaacacgt gttagatgcc cttgacccca atgcttatga agccttcaaa
     1741 gcatcgcgca acctgcagga tgtggtggag cgagtccttc acaatgagca ggaggtggag
     1801 aaggacccag gccggcgacg ggctctgagg gtccaggcct cactgatgac gcctgtgcag
     1861 cccatgctgg ctgaggcctg caaatccatt gaatatgcaa tgaagaagtg tcccaatggc
     1921 atgttctctg agatcaagta cgatggtgag cgagtccagg tgcataagaa gggggaccac
     1981 ttcagctact tcagccgcag tctcaagcct gtcctgcctc acaaggtggc ccacttcaag
     2041 gactatatac ccaaagcttt tcctgggggt caaagcatga tcttggattc cgaggtactc
     2101 ctgattgaca acaacacagg caaaccactg ccctttggga ctctgggagt gcacaagaaa
     2161 gctgctttcc aggatgctaa tgtctgcctg tttgtttttg attgtatcta ctttaatgat
     2221 gtcagcttga tggacaggcc tctctgtgag cgacggaagt ttcttcatga caacatggtt
     2281 gaaatccgta accggatcat gttctcagaa atgaagcaag tcacaaaagc ttcagacctg
     2341 gctgacatga taaaccgggt gatccgagag ggcttggaag ggttggtgct aaaggatgtg
     2401 aagggtacat atgagcctgg gaagcggcac tggctgaaag ttaagaaaga ttatttgaat
     2461 gagggggcca tggctgatac agctgatctg gtggttcttg gggccttcta tgggcaaggg
     2521 agcaaaggcg gaatgatgtc catcttcctc atgggctgct atgaccctga cagccaaaag
     2581 tggtgcactg ttacaaaatg tgctggaggc catgatgatg ccacacttgc ccgcctgcag
     2641 aaagagctgg ttatggtgaa gatcagcaag gatcctagta agatacccag ctggctgaag
     2701 atcaacaaga tctattatcc tgactttatc gtcccagacc caaagaaagc tgctgtgtgg
     2761 gagatcacag gggcagaatt ctccagatct gaggcacaca ctgctgatgg catctccatc
     2821 cgatttcctc gatgcacccg aatccgggac gacaaggact ggaaatctgc cactaacctc
     2881 ccccaactca aggaactgta ccagctgtcc aaagacaagg cagactttgc tgtggtggct
     2941 ggagatgagg caagccccac tactggaggc agcagtggcg agaacgaggg cactgctggg
     3001 tctgctgggc cctgcaaggg cccccccagt aagtcctctg ccagtgccaa gaccaccgaa
     3061 cagaagctca atagccccag tagcagaggt ggcatcaaac cgattccaaa gcattccccc
     3121 atgaaaccag gagagaagct ggctgtgaag tcttctccag tcaaagtggg aatgaagagg
     3181 aaagctactg atgagacccc atgcctgaaa aagtaataat ctagagggcc cgtttaaacc
     3241 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3301 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3361 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3421 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3481 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3541 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3601 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3661 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3721 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3781 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3841 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3901 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3961 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4141 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4201 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4261 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4321 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4381 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4441 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4501 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4561 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4621 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4681 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4741 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4801 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4861 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4921 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4981 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5041 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5101 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5161 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5221 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5281 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5461 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5521 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5581 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5641 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5701 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6421 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6481 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6541 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6601 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6661 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6721 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6781 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6841 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6901 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6961 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7021 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7081 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7141 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7201 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7261 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7321 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7381 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7441 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7501 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7561 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7621 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7681 cccctatggt gcactctcag tacaatctgc tctgatg
//
