LOCUS       62056_23245        7316 bp DNA     circular SYN 21-MAR-2021
DEFINITION  Expression vector RFP, complete sequence.
ACCESSION   MW560964
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  AUTHORS   Park SB.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-FEB-2021) NIDDK, NIH, 10 Center Drive, Bethesda, MD 
            20814, USA
REFERENCE   2  (bases 1 to 7316)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (01-FEB-2021) NIDDK, NIH, 10 Center Drive, Bethesda, MD 20814, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(968..1825)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1826..1930)
                     /label=AmpR promoter
     rep_origin      complement(1956..2411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2553..2569
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2579..2597
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2630..3808
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3821..4417
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     enhancer        4517..4896
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4897..5100
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          5212..5784
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             5851..6543
                     /codon_start=1
                     /label=TurboRFP
                     /note="red fluorescent protein from Entacmaea quadricolor"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     polyA_signal    6707..6762
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7120..7136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7144..7160)
                     /label=lac operator
                     /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(7168..7198)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7213..7234)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     1981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2401 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2461 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2521 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     2581 atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgcg gctccggtgc
     2641 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2701 caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2761 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2821 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     2881 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     2941 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     3001 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     3061 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     3121 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     3181 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     3241 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     3301 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     3361 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     3421 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     3481 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     3541 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     3601 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     3661 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     3721 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     3781 tttttttctt ccatttcagg tgtcgtgacg tacggccacc atgaccgagt acaagcccac
     3841 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     3901 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     3961 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4021 ggacgacggc gccgccgtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4081 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4141 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4201 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4261 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4321 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4381 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gcggccgctc tagagagctt
     4441 ggcccattgc atacgttgta tccatatcat aatatgtaca tttatattgg ctcatgtcca
     4501 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     4561 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4621 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4681 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4741 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     4801 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     4861 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     4921 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     4981 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     5041 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     5101 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     5161 agacaccggg accgatccag cctccggtcg accgatcctg agaacttcag ggtgagtttg
     5221 gggacccttg attgttcttt ctttttcgct attgtaaaat tcatgttata tggagggggc
     5281 aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca tggaccctca
     5341 tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc tcttattttc
     5401 ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc atttgtaacg aatttttaaa
     5461 ttcacttttg tttatttgtc agattgtaag tactttctct aatcactttt ttttcaaggc
     5521 aatcagggta tattatattg tacttcagca cagttttaga gaacaattgt tataattaaa
     5581 tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat tcttattggt
     5641 agaaacaact acaccctggt catcatcctg cctttctctt tatggttaca atgatataca
     5701 ctgtttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc taaccatgtt
     5761 catgccttct tctctttcct acagctcctg ggcaacgtgc tggttgttgt gctgtctcat
     5821 cattttggca aagaattcct cgacggatcc atgagcgagc tgatcaagga gaacatgcac
     5881 atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc
     5941 gaaggcaagc cctacgaggg cacccagacc atgaagatca aggtggtcga gggcggccct
     6001 ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa agccttcatc
     6061 aaccacaccc agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg
     6121 gagagaatca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcttc
     6181 cagaacggct gcatcatcta caacgtcaag atcaacgggg tgaacttccc atccaacggc
     6241 cctgtgatgc agaagaaaac acgcggctgg gaggccaaca ccgagatgct gtaccccgct
     6301 gacggcggcc tgagaggcca cagccagatg gccctgaagc tcgtgggcgg gggctacctg
     6361 cactgctcct tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc
     6421 ggcttccact tcgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac
     6481 gtcgagcagc acgagatggc tgtggccaag tactgcgacc tccctagcaa actggggcac
     6541 agatgaggat ccgtcgagga attcactcct caggtgcagg ctgcctatca gaaggtggtg
     6601 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     6661 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     6721 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     6781 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     6841 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     6901 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     6961 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     7021 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     7081 tggagatccc tcgacggatc ggccgcaatt cgtaatcatg tcatagctgt ttcctgtgtg
     7141 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7201 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7261 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cgggga
//
