LOCUS       Exported                7215 bp ds-DNA     circular SYN 19-JAN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 19, 2019 from SnapGene Viewer 4.3.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7215
                     /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             913..1542
                     /codon_start=1
                     /label=Rab 2 OFF
                     /note="Vector contains Rab2 OFF mutation, BirA* and MAO."
                     /translation="MAYAYLFKYIIIGDTGVGKNCLLLQFTDKRFQPVHDLTIGVEFGA
                     RMITIDGKQIKLQIWDTAGQESFRSITRSYYRGAAGALLVYDITRRDTFNHLTTWLEDA
                     RQHSNSNMVIMLIGNKSDLESRREVKKEEGEAFAREHGLIFMETSAKTASNVEEAFINT
                     AKEIYEKIQEGVFDINNEANGIKIGPQHAATNATHAGNQGGQQAGGG"
     CDS             2563..2589
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2815..3039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3085..3513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3527..3856
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3707..3842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3923..4717
                     /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    4891..5012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5061..5077)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5085..5101
                     /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(5109..5139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5154..5175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5463..6048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6219..7079)
                     /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(7080..7184)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
      901 aagcttaccg gtatggcgta cgcctatctc ttcaagtaca tcataatcgg cgacacaggt
      961 gttggtaaaa actgcttatt gctacagttt acagacaaga ggtttcagcc agtgcatgac
     1021 cttactattg gtgtagagtt cggtgctcga atgataacta ttgatgggaa acagataaaa
     1081 cttcagatat gggatacggc agggcaagaa tcctttcgtt ccatcacaag gtcgtattac
     1141 agaggtgcag caggagcttt actagtttac gatattacac ggagagatac attcaaccac
     1201 ttgacaacct ggttagaaga tgcccgccag cattccaatt ccaacatggt cattatgctt
     1261 attggaaata aaagtgattt agaatctaga agagaagtaa aaaaagaaga aggtgaagct
     1321 tttgcacgag aacatggact catcttcatg gaaacgtctg ctaagactgc ttccaatgta
     1381 gaagaggcat ttattaatac agcaaaagaa atttatgaaa aaattcaaga aggagtcttt
     1441 gacattaata atgaggcaaa tggcattaaa attggccctc agcatgctgc taccaatgca
     1501 acacatgcag gcaatcaggg aggacagcag gctgggggcg gcgaattcgg agccggagct
     1561 ggtaccatgg acaaggacaa caccgtgccc ctgaagctga tcgccctgct ggccaacggc
     1621 gagttccact ctggcgagca gctgggagag accctgggaa tgagcagagc cgccatcaac
     1681 aagcacatcc agacactgag agactgggga gtggacgtgt tcaccgtgcc tggcaagggc
     1741 tacagcctgc ctgagcctat ccagctgctg aacgccaagc agatcctggg acagctggat
     1801 ggcggaagcg tggccgtgct gcctgtgatc gactccacca atcagtacct gctggacaga
     1861 atcggagagc tgaagtccgg cgacgcctgc atcgccgagt accagcaggc tggcagagga
     1921 ggcagaggac ggaagtggtt cagcccattc ggagccaacc tgtacctgtc catgttctgg
     1981 agactggagc agggacctgc tgctgccatc ggactgagtc tggtgatcgg aatcgtgatg
     2041 gccgaggtgc tgagaaagct gggagccgac aaggtgagag tgaagtggcc taatgacctg
     2101 tacctccagg accgcaagct ggctggcatc ctggtggagc tgacaggcaa gacaggcgat
     2161 gccgctcaga tcgtgatcgg agccggaatc aacatggcca tgagaagagt ggaggagagc
     2221 gtggtgaacc agggctggat caccctgcag gaggctggca tcaacctgga ccggaacacc
     2281 ctggccgcca tgctgatcag agagctgaga gccgctctgg agctgttcga gcaggaggga
     2341 ctggctcctt acctgagcag atgggagaag ctggacaact tcatcaacag acctgtgaag
     2401 ctgatcatcg gcgacaagga aatcttcggc atctccagag gaatcgacaa gcagggagct
     2461 ctgctgctgg agcaggacgg aatcatcaag ccctggatgg gcggagaaat ctccctgaga
     2521 agcgcagaga agggatccgt cgacggagcc ggtgctggcg cataccccta cgacgtgccc
     2581 gactacgcag gctccgcagt tccagcggta gaaatcaccc acaccttctg ggaaaggaac
     2641 ctgccctctg tttctggcct gctgaagatc attggatttt ccacatcagt aactgccctg
     2701 gggtttgtgc tgtacaaata caagctcctg ccacggtctt gacccgggga tatccagcac
     2761 agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     2821 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2881 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2941 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3001 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3061 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3121 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3181 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3241 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3301 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3361 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3421 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3481 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3601 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3721 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3781 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3841 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3901 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3961 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4021 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4081 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4141 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4201 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4261 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4321 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4381 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4441 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4501 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4561 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4621 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4681 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4741 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4801 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4861 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4921 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4981 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5041 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5101 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5161 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5221 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5281 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5341 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5401 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5881 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7201 gtgccacctg acgtc
//
