LOCUS       Exported                7985 bp DNA     circular SYN 14-JAN-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7985)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 14, 2022 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    769..789
                     /locus_tag="CMV F"
                     /label=CMV F
                     /note="color: #00ffff"
     primer_bind     863..882
                     /locus_tag="T7"
                     /label=T7
                     /note="color: #ff8080; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    944..3279
                     /locus_tag="mTfR1 (Shall Jue)"
                     /label=mTfR1 (Shall Jue)
                     /note="color: #eeeeee"
     misc_feature    991..3279
                     /locus_tag="TfR1 ORF (JD)"
                     /label=TfR1 ORF (JD)
                     /note="color: #ffff80"
     misc_feature    2919..2936
                     /locus_tag="TfR1 internal forward"
                     /label=TfR1 internal forward
                     /note="/ApEinfo_hidden"
                     /note="color: #00ffff"
     misc_feature    3280..3297
                     /locus_tag="6 x GS linker"
                     /label=6 x GS linker
                     /note="color: #c0c0c0"
     misc_feature    3298..3303
                     /locus_tag="AgeI site"
                     /label=AgeI site
                     /note="color: #ff9e2b"
     misc_feature    3304..3423
                     /locus_tag="miniR"
                     /label=miniR
                     /note="color: #ff80ff"
     misc_feature    3424..3429
                     /locus_tag="2nd AgeI site"
                     /label=2nd AgeI site
                     /note="color: #ff5219"
     misc_feature    3430..3432
                     /locus_tag="TfR1 ORF "
                     /label=TfR1 ORF
                     /note="color: #ffff80"
     promoter        complement(3537..3555)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    3575..3592
                     /locus_tag="BGH rev"
                     /label=BGH rev
                     /note="color: #00ffff"
     polyA_signal    3581..3805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3851..4279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4293..4623
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4474..4609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4690..5484
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4693..5484
                     /locus_tag="Kan/NeoR"
                     /label=Kan/NeoR
                     /note="color: #d5d5ff"
     polyA_signal    5658..5779
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5828..5844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5852..5868
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5876..5906)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5876 .. 5882 / #ffffff / -10
                        2: 5883 .. 5900 / #ffffff
                        3: 5901 .. 5906 / #ffffff / -35"
     protein_bind    5921..5942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     misc_feature    6212..6894
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
                     /note="color: #c0c0c0"
     rep_origin      complement(6230..6818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6989..7849)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6989 .. 7780 / #ccffcc
                        2: 7781 .. 7849 / #ccffcc / signal sequence
                     Cleavage site after base 7780"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    6992..7651
                     /locus_tag="AmpR"
                     /label=AmpR
                     /note="color: #ffdddd"
     promoter        complement(7850..7954)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
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 ccactagtaa cggccgccag tgtgctggaa ttccgcggac tgtagaggcg
      961 cttcctagta ctcccttgta gcagctgaga atgatggatc aagccagatc agcattctct
     1021 aacttgtttg gtggggaacc attgtcatac acccggttta gccttgctcg gcaagtagat
     1081 ggagataaca gtcatgtgga gatgaaactg gctgcagatg aagaagaaaa tgccgacaat
     1141 aacatgaagg ctagtgtcag aaaacccaag aggtttaatg gaagactctg ctttgcagct
     1201 attgcactag tcattttctt cttgattgga ttcatgagtg gctacctggg ctattgtaag
     1261 cgtgtagaac aaaaagagga gtgtgtgaaa ctggctgaaa cggaggagac agacaagtca
     1321 gaaaccatgg aaacagagga tgttcctaca tcatctcgct tatattgggc agacctcaaa
     1381 acactgttgt cagagaagtt gaactccata gagtttgctg acaccatcaa gcagctgagc
     1441 cagaatacat acactcctcg tgaggctgga tctcaaaaag atgaaagtct tgcctattat
     1501 attgaaaatc agttccatga atttaaattc agcaaagtct ggcgagatga acactatgtg
     1561 aagattcaag tgaaaagcag cattggtcaa aacatggtga ccatagtgca gtcaaatggt
     1621 aacttagacc cagtggagtc tcccgagggt tatgtggcat tcagtaaacc tacagaagtt
     1681 tctggtaaac tggtccatgc taattttggc actaaaaagg actttgaaga actaagttat
     1741 tctgtgaatg gatctttagt gattgttaga gcaggggaaa ttacttttgc agaaaaggtt
     1801 gcaaatgccc aaagctttaa tgcaattggt gtcctcatat acatggacaa gaataaattc
     1861 cccgttgttg aggcagacct tgcactcttt ggacatgctc atctaggaac tggtgatcca
     1921 tacacacctg gctttccttc tttcaatcat actcagtttc cgccatctca gtcatcaggg
     1981 ttgcctaata tacctgtgca aacaatctca agagctgctg cagaaaagct atttggaaaa
     2041 atggaaggaa gctgtcctgc tagatggaac atagattctt catgtaagct ggaactttca
     2101 cagaatcaaa atgtgaagct cattgtgaaa aacgtactga aagaaagaag aatacttaac
     2161 atctttggag ttattaaagg ttatgaggaa ccagaccgtt atgttgtagt aggagcccag
     2221 agagacgctt tgggtgctgg tgttgcggcg aagtccagtg tgggaacagg tcttctgttg
     2281 aaacttgccc aagtattctc agatatgatt tcaaaagatg gatttagacc cagcagaagt
     2341 ataatctttg ccagctggac tgcaggcgac tttggagctg ttggtgccac tgagtggttg
     2401 gagggatacc tttcatcttt gcatttaaaa gctttcactt atattaattt ggataaagtt
     2461 gtccttggta ctagtaactt caaagtttct gccagcccct tattatatac acttatggga
     2521 aagataatgc aagatgtaaa gcatccagtt gatggaaaat ctctatatag agacagcaat
     2581 tggattagca aagttgagaa actttccttt gacaatgctg catatccttt ccttgcatat
     2641 tctggaatcc cagcagtttc tttttgtttt tgtgaggatg cagactatcc ttatttgggc
     2701 actagattgg atacctatga ggcattgact cagaaagttc ctcagctcaa ccaaatggtt
     2761 cgtacagcag cggaagtggc tggtcagctc attattaaac ttacccatga cgttgaattg
     2821 aacctggact atgagatgta taacagcaaa ctactgtcat ttatgaagga tctgaaccag
     2881 ttcaaaacag atatcaggga tatgggtcta agtctacagt ggctgtattc cgctcgtgga
     2941 gactacttcc gtgctacttc tagactaaca actgattttc ataatgctga gaaaacaaac
     3001 agatttgtca tgagggaaat caatgatcgt attatgaaag tggagtatca cttcctgtcg
     3061 ccctatgtat ctccaagaga gtctcctttc cgacatatct tctggggctc tggctctcac
     3121 actctctcag ctttagtgga gaacttgaag cttcgtcaaa aaaatattac tgcttttaat
     3181 gaaaccctct tcagaaacca gttggccctg gctacttgga ctattcaggg agtcgcaaat
     3241 gccctctctg gtgacatttg gaatattgac aatgagtttg gtagcggcag cggtagcacc
     3301 ggtatgggtt cttctctcga aatccgtgtt gcgttcctgc gtcagcgtaa caccgcgctg
     3361 cgtaccgaag ttgcggaact ggaacaggaa gttcagcgtc tggaaaaccg ttacggtccg
     3421 ctgaccggtt aaatgtaaca tgcataatta aataagagca gggtagtctg tttctagaat
     3481 tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg ccctattcta
     3541 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3601 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3661 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3721 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3781 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3841 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3901 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3961 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     4021 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4081 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4141 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4201 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     4261 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4321 caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4441 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4501 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4561 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4621 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4681 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4741 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4801 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4861 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4921 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4981 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5041 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5101 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5161 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5221 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5281 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5341 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5401 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5461 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5521 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5581 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5641 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5701 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5761 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5821 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5881 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5941 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6001 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6061 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6421 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6661 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7981 acgtc
//
