LOCUS       Exported                6097 bp ds-DNA     circular SYN 30-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDuex2-rsTagRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6097)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 12, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6097
                     /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        858..886
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    940..945
                     /label=XhoI
     CDS             946..1656
                     /codon_start=1
                     /product="TagRFP derivative with reversibly photoswitchable
                     absorbance (Subach et al., 2010)"
                     /label=rsTagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEAATEMLYPADGGLEGRGD
                     MALKLVGGGHLICNLKTTYRSKNPAKNLKMPGVYFVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    1660..1665
                     /label=XbaI
     polyA_signal    1697..1921
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1967..2395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2409..2738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2589..2724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2805..3599
                     /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    3773..3894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3943..3959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3967..3983
                     /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(3991..4021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4036..4057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4345..4930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5101..5961)
                     /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(5962..6066)
                     /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 gtccggattg acaattaatc atcggctcgt ataatatacc cgtttttttg
      901 ggctagaaat aattttgtct aactttaaga aggagatatc tcgagatggt gtctaagggc
      961 gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtgaacaac
     1021 caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
     1081 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc
     1141 ttcatgtacg gcagccgcac cttcatcaac cacacccagg gcatccccga cttctggaag
     1201 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
     1261 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagctc
     1321 agaggggtga acttcccatc caacggccct gtgatgcaga agaaaacact cggctgggag
     1381 gccgccaccg agatgctgta ccccgctgac ggcggcctgg aaggcagagg ggacatggcc
     1441 ctgaagctcg tgggcggggg ccacctgatc tgcaacttga agaccacata cagatccaag
     1501 aatcccgcta agaacctcaa gatgcccggc gtctactttg tggaccacag actggaaaga
     1561 atcaaggagg ccgacaaaga gacctacgtc gagcagcacg aggtggctgt ggccagatac
     1621 tgcgacctcc ctagcaaact ggggcacaag cttaattaat ctagagggcc cgtttaaacc
     1681 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1741 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1801 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1861 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1921 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1981 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2041 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2101 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2161 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2221 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2281 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2341 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2401 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2461 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2521 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2581 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2641 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2701 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2761 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2821 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2881 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2941 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3001 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3061 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3121 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3181 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3241 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3301 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3361 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3421 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3481 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3541 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3601 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3661 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3721 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     3781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3901 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     3961 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4021 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4081 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4141 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4201 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4861 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4921 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4981 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5041 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5221 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5341 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5401 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5461 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5521 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5581 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5641 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5701 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5761 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5821 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5881 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5941 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6001 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6061 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
