LOCUS       sequence-381564-        6588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split TEV assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6588)
  AUTHORS   Wu Y, von Hauff IV, Jensen N, Rossner MJ, Wehr MC
  TITLE     Improved Split TEV GPCR beta-arrestin-2 Recruitment Assays via 
            Systematic Analysis of Signal Peptide and beta-arrestin Binding 
            Motif Variants.
  JOURNAL   Biosensors (Basel). 2022 Dec 29;13(1):48. doi: 10.3390/bios13010048.
  PUBMED    36671883
REFERENCE   2  (bases 1 to 6588)
  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: "Biosensors 
            (Basel)."; date: "2022-12-29"; volume: "13(1):48. doi"; pages: " 
            10.3390/bios13010048"
COMMENT     Sequence Label: pcDNA3.1_Zeo_ARRB2-1-383-CTEV-2xHA
FEATURES             Location/Qualifiers
     source          1..6588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    941..965
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2118..2142)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2492..2518
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2522..2548
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2596..2613)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2602..2826
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2872..3300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2959..2978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3169..3190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3309..3329)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3314..3643
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3494..3629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3556..3575
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3691..3738
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3757..4131
                     /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    4261..4382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4298..4317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4352..4371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4431..4447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4431..4447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4444..4466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4455..4471
                     /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(4479..4485,4486..4503,4504..4509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4524..4545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4662..4679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4833..5421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4913..4932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5592..6383,6384..6452))
                     /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     6215..6234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6453..6557)
                     /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 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgcgatatca acaagtttgt acaaaaaagc
      961 aggctctacc atgggggaga aacccgggac cagggtcttc aagaagtcga gccctaactg
     1021 caagctcacc gtgtacttgg gcaagcggga cttcgtagat cacctggaca aagtggaccc
     1081 tgtagatggc gtggtgcttg tggaccctga ctacctgaag gaccgcaaag tgtttgtgac
     1141 cctcacctgc gccttccgct atggccgtga agacctggat gtgctgggct tgtccttccg
     1201 caaagacctg ttcatcgcca cctaccaggc cttccccccg gtgcccaacc caccccggcc
     1261 ccccacccgc ctgcaggacc ggctgctgag gaagctgggc cagcatgccc accccttctt
     1321 cttcaccata ccccagaatc ttccatgctc cgtcacactg cagccaggcc cagaggatac
     1381 aggaaaggcc tgcggcgtag actttgagat tcgagccttc tgtgctaaat cactagaaga
     1441 gaaaagccac aaaaggaact ctgtgcggct ggtgatccga aaggtgcagt tcgccccgga
     1501 gaaacccggc ccccagcctt cagccgaaac cacacgccac ttcctcatgt ctgaccggtc
     1561 cctgcacctc gaggcttccc tggacaagga gctgtactac catggggagc ccctcaatgt
     1621 aaatgtccac gtcaccaaca actccaccaa gaccgtcaag aagatcaaag tctctgtgag
     1681 acagtacgcc gacatctgcc tcttcagcac cgcccagtac aagtgtcctg tggctcaact
     1741 cgaacaagat gaccaggtat ctcccagctc cacattctgt aaggtgtaca ccataacccc
     1801 actgctcagt gacaaccggg agaagcgggg tctcgccctg gatgggaaac tcaagcacga
     1861 ggacaccaac ctggcttcca gcaccatcgt gaaggagggt gccaacaagg aggtgctggg
     1921 aatcctggtg tcctacaggg tcaaggtgaa gctggtggtg tctcgaggcg gggatgtctc
     1981 tgtggagctg ccttttgttc ttatgcaccc caagccccac gaccacatcc ccctccccag
     2041 accccagtca gccgctccgg agacagatgt ccctgtggac accaacctca ttgaatttga
     2101 taccaactat gccacagacc cagctttctt gtacaaagtg gttgatatcg gaggtggagg
     2161 tagcggaggt ggaggtagca agagcatgtc tagcatggtg tcagacacta gctgcacatt
     2221 cccttcatct gatggtatat tctggaagca ttggattcaa accaaggatg ggcagtgtgg
     2281 cagtccatta gtatcaacta gagatgggtt cattgttggt atacactcag catcgaattt
     2341 caccaacaca aacaattatt tcacaagcgt gccgaaaaac ttcatggaat tgttgacaaa
     2401 tcaggaggcg cagcagtggg ttagtggttg gcgattaaac gctgactcag tattgtgggg
     2461 gggccataaa gttttcatgc ccaaacctgg ctacccatac gatgttcctg actatgcggg
     2521 ctatccctat gacgtcccgg actatgcatg aggatccgag ctcggtacca agcttaagtt
     2581 taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2641 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2701 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2761 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2821 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2881 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2941 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3001 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3061 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3121 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3181 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3241 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3301 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3481 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3601 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3661 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     3721 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     3781 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     3841 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     3901 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     3961 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     4021 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     4081 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     4141 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4201 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4261 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4321 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4381 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4441 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4501 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4561 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4621 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5461 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5521 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5581 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5641 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5701 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5761 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5821 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5881 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5941 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6001 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6061 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6121 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6181 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6241 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6301 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6361 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6421 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6481 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6541 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
