LOCUS       sequence-240794-        6991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Violin JD, Ren XR, Lefkowitz RJ
  TITLE     G-protein-coupled receptor kinase specificity for beta-arrestin 
            recruitment to the beta2-adrenergic receptor revealed by 
            fluorescence resonance energy transfer.
  JOURNAL   J Biol Chem. 2006 Jul 21;281(29):20577-88. Epub 2006 May 10.
  PUBMED    16687412
REFERENCE   2  (bases 1 to 6991)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2006-07-21"; volume: "281(29)"; pages: "20577-88. 
            Epub 2006 May 10"
COMMENT     Sequence Label: beta-arrestin1 mYFP
FEATURES             Location/Qualifiers
     source          1..6991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        463..666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     617..637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     711..730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        711..729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2099..2101,2102..2815)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2141..2162)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2402..2421)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2749..2770
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2847..2864)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2853..3077
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3123..3551
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3210..3229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3420..3441
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3560..3580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3565..3894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3745..3880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3807..3826
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3942..3989
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4008..4382
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4512..4633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4549..4568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4603..4622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4682..4698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4682..4698)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4695..4717)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4706..4722
                     /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(join(4730..4736,4737..4754,4755..4760))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4775..4796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4913..4930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5084..5672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5164..5183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5843..6634,6635..6703))
                     /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     6466..6485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6704..6808)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6883..6902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
       61 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      121 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      181 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      421 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      481 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      541 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      601 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      661 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      721 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc gagctcggat
      781 ccactagtaa cggccgccag tgtgctggaa ttctgcagat cggaccatgg gcgacaaagg
      841 gacacgagtg ttcaagaagg caagccccaa tggaaagctc accgtctacc tgggaaagcg
      901 ggactttgtg gaccacattg acctggtgga ccccgtggat ggcgtggtcc tggtggatcc
      961 tgagtatctc aaagaaaggc gagtctacgt gacactgacc tgcgccttcc ggtatggccg
     1021 ggaagacctg gatgtcttgg gtctgacttt tcgcaaagac ctgtttgtgg ctaacgtgca
     1081 gtccttccca ccggcccctg aggacaagaa gccactgact cggctacaag agcgactcat
     1141 caagaagctg ggcgagcatg cctacccctt cacctttgag atcccgccaa accttccgtg
     1201 ctcagtcaca ttgcaacctg ggcctgagga cacagggaag gcctgcggtg tggattatga
     1261 agtgaaagcc ttctgtgctg agaacctgga ggagaagatc cacaaaagga attctgtgcg
     1321 gctagtcatc cggaaggttc aatatgcccc tgagaggcct ggccctcagc ccacggctga
     1381 gaccaccaga cagttcctca tgtcggacaa gcccctgcac cttgaggcat ctctggataa
     1441 ggagatctat tatcatggag aacccatcag cgtcaatgtc catgtcacca acaacaccaa
     1501 caagactgtg aagaagatca agatctcggt gcgccagtat gcagacatct gtctcttcaa
     1561 cacagctcag tacaagtgcc cagtggccat ggaggaagct gatgatactg tggcacccag
     1621 ctcaacattc tgcaaggtct acacactgac tcccttcctg gcaaacaaca gagagaagcg
     1681 ggggcttgcc ctcgacggga agctcaagca tgaagacaca aatctggctt ccagcactct
     1741 gttgcgggaa ggcgccaacc gtgaaatcct gggtatcatt gtttcctaca aagtcaaagt
     1801 gaagctggtg gtgtcccggg gcggcctgtt gggagacctt gcatccagtg acgtggctgt
     1861 ggaactgccc tttaccttaa tgcaccccaa gcctaaagag gagcccccac atcgggaagt
     1921 tccagagagc gagactccag tagacaccaa tctcatagag cttgacacca atgatgacga
     1981 cattgtgttt gaggactttg ctcgtcagcg gctgaaaggc atgaaggatg acaaggacga
     2041 agaggatgat ggcaccggct ctccacacct caacaacaga ggtggactcg aggcactggt
     2101 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2161 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2221 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2281 gaccaccttc ggctacggcc tgaagtgctt cgcccgctac cccgaccaca tgaagcagca
     2341 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2401 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2461 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2521 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2581 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2641 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2701 gagctaccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2761 ggagttcgtg accgccgccg ggatcactca cggcatggac gagctgtaca agtaatctag
     2821 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3121 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3181 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3241 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3301 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3361 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3421 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3481 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3541 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3901 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     3961 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     4021 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     4081 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     4141 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     4201 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     4261 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     4321 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     4381 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4441 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4501 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4621 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4681 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4741 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4801 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4861 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4921 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4981 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5041 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5101 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5161 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5221 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5281 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5341 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5401 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5461 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5521 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5581 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5641 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5701 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5761 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5821 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5881 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5941 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6001 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6061 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6121 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6181 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6241 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6301 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6361 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6421 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6481 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6541 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6601 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6661 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     6841 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     6901 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     6961 gagcaaaatt taagctacaa caaggcaagg c
//
