LOCUS       Exported                7079 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Beta-2-adrenergic receptor internally tagged with miRFP670nano.
ACCESSION   .
VERSION     .
KEYWORDS    pB2AR-miRFP670nano
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 7079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7079
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        176..555
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        556..759
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     710..730
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     804..823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        804..822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2611..2628)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2617..2841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2887..3315
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2974..2993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3324..3344)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3329..3658
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3509..3644
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3571..3590
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3725..4519
                     /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"
     primer_bind     complement(3779..3798)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4389..4408
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4693..4814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4730..4749)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4784..4803
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4863..4879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4863..4879)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4876..4898)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4887..4903
                     /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(4911..4941)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4956..4977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5094..5111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5265..5853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5345..5364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6024..6884)
                     /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"
     primer_bind     6647..6666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6885..6989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7064..4)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
       61 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      121 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      181 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      241 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      301 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      361 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      421 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      481 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      541 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      601 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      661 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      721 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      781 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
      841 catggggcaa cccgggaacg gcagcgcctt cttgctggca cccaatagaa gccatgcgcc
      901 ggaccacgac gtcacgcagc aaagggacga ggtgtgggtg gtgggcatgg gcatcgtcat
      961 gtctctcatc gtcctggcca tcgtgtttgg caatgtgctg gtcatcacag ccattgccaa
     1021 gttcgagcgt ctgcagacgg tcaccaacta cttcatcact tcactggcct gtgctgatct
     1081 ggtcatgggc ctggcagtgg tgccctttgg ggccgcccat attcttatga aaatgtggac
     1141 ttttggcaac ttctggtgcg agttttggac ttccattgat gtgctgtgcg tcacggccag
     1201 cattgagacc ctgtgcgtga tcgcagtgga tcgctacttt gccattactt cacctttcaa
     1261 gtaccagagc ctgctgacca agaataaggc ccgggtgatc attctgatgg tgtggattgt
     1321 gtcaggcctt acctccttct tgcccattca gatgcactgg taccgggcca cccaccagga
     1381 agccatcaac tgctatgcca atgagacctg ctgtgacttc ttcacgaacc aagcctatgc
     1441 cattgcctct tccatcgtgt ccttctacgt tcccctggtg atcatggtct tcgtctactc
     1501 cagggtcttt caggaggcca aaaggcagct ccagaagatt gacaaatctg agggccgctt
     1561 ccatgtccag aaccttagcc aggtggaggg atccggtggg ggaggcagca tggcaaacct
     1621 ggacaagatg ctgaatacca cagtaacaga ggtgcggcag ttcctgcagg tggacagagt
     1681 gtgcgtgttc cagtttgagg aggattatag cggagtggtg gtggtggagg ccgtggacga
     1741 taggtggatc tccatcctga agacccaggt gcgggataga tacttcatgg agacaagggg
     1801 cgaggagtat tctcacggcc gctaccaggc catcgccgac atctacaccg caaacctgac
     1861 agagtgctac agggatctgc tgacacagtt tcaggtgaga gcaatcctgg ccgtgcccat
     1921 cctgcagggc aagaagctgt ggggcctgtt ggtggcacac cagctggcgg cccctagaca
     1981 gtggcagacc tgggagatcg actttctgaa gcagcaggcc gtggtggtgg gcatcgccat
     2041 ccagcagagc ggaggtggcg ggagcaccgg tcaggatggg cggacggggc atggactccg
     2101 cagatcttcc aagttctgct tgaaggagca caaagccctc aagacgttag gcatcatcat
     2161 gggcactttc accctctgct ggctgccctt cttcatcgtt aacattgtgc atgtgatcca
     2221 ggataacctc atccgtaagg aagtttacat cctcctaaat tggataggct atgtcaattc
     2281 tggtttcaat ccccttatct actgccggag cccagatttc aggattgcct tccaggagct
     2341 tctgtgcctg cgcaggtctt ctttgaaggc ctatgggaat ggctactcca gcaacggcaa
     2401 cacaggggag cagagtggat atcacgtgga acaggagaaa gaaaataaac tgctgtgtga
     2461 agacctccca ggcacggaag actttgtggg ccatcaaggt actgtgccta gcgataacat
     2521 tgattcacaa gggaggaatt gtagtacaaa tgactcactg ctgtaagcgg ccgctcgagt
     2581 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2641 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2701 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2761 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2821 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2881 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3301 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3361 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3481 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3541 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3601 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3661 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3721 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3781 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3841 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3901 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3961 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4021 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4081 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4141 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4201 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4261 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4321 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4381 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4441 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4501 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4561 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4621 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4681 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4741 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4801 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7021 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgat
//
