LOCUS       sequence-456016-        7030 bp    DNA     circular SYN 15-APR-2025
DEFINITION  ADRA1A receptor for split TEV assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7030)
  AUTHORS   Popovic L, Brankatschk B, Palladino G, Rossner MJ, Wehr MC
  TITLE     Ischemic stroke alleviating Tat-N peptide AVLX-144 ameliorates 
            abnormal molecular signatures in a cellular model of ERBB4 
            deficiency
  JOURNAL   In preparation
  PUBMED    39405910
REFERENCE   2  (bases 1 to 7030)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "In 
            preparation"
COMMENT     Sequence Label: pTag4C_ADRA1A-V2R-NTEV-tcs-GV
FEATURES             Location/Qualifiers
     source          1..7030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    632..656
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2088..2112)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2603..2623
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2630..2659
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2669..3109
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             3152..3172
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3323..3556
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     regulatory      3563..3572
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3575..3601
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3818..3939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3855..3874)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3909..3928
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3946..4401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4083..4104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4295..4314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4428..4532
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4534..4891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4534..4725
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4557..4577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4742..4877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4926..5720
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4926..5720
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4980..4999)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5590..5609
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5908..5927)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5952..5999
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6328..6916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6817..6836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatc aacaagtttg tacaaaaaag caggctctac
      661 catggtgttt ctctcgggaa atgcttccga cagctccaac tgcacccaac cgccggcacc
      721 ggtgaacatt tccaaggcca ttctgctcgg ggtgatcttg gggggcctca ttcttttcgg
      781 ggtgctgggt aacatcctag tgatcctctc cgtagcctgt caccgacacc tgcactcagt
      841 cacgcactac tacatcgtca acctggcggt ggccgacctc ctgctcacct ccacggtgct
      901 gcccttctcc gccatcttcg aggtcctagg ctactgggcc ttcggcaggg tcttctgcaa
      961 catctgggcg gcagtggatg tgctgtgctg caccgcgtcc atcatgggcc tctgcatcat
     1021 ctccatcgac cgctacatcg gcgtgagcta cccgctgcgc tacccaacca tcgtcaccca
     1081 gaggaggggt ctcatggctc tgctctgcgt ctgggcactc tccctggtca tatccattgg
     1141 acccctgttc ggctggaggc agccggcccc cgaggacgag accatctgcc agatcaacga
     1201 ggagccgggc tacgtgctct tctcagcgct gggctccttc tacctgcctc tggccatcat
     1261 cctggtcatg tactgccgcg tctacgtggt ggccaagagg gagagccggg gcctcaagtc
     1321 tggcctcaag accgacaagt cggactcgga gcaagtgacg ctccgcatcc atcggaaaaa
     1381 cgccccggca ggaggcagcg ggatggccag cgccaagacc aagacgcact tctcagtgag
     1441 gctcctcaag ttctcccggg agaagaaagc ggccaaaacg ctgggcatcg tggtcggctg
     1501 cttcgtcctc tgctggctgc cttttttctt agtcatgccc attgggtctt tcttccctga
     1561 tttcaagccc tctgaaacag tttttaaaat agtattttgg ctcggatatc taaacagctg
     1621 catcaacccc atcatatacc catgctccag ccaagagttc aaaaaggcct ttcagaatgt
     1681 cttgagaatc cagtgtctct gcagaaagca gtcttccaaa catgccctgg gctacaccct
     1741 gcacccgccc agccaggccg tggaagggca acacaaggac atggtgcgca tccccgtggg
     1801 atcaagagag accttctaca ggatctccaa gacggatggc gtttgtgaat ggaaattttt
     1861 ctcttccatg ccccgtggat ctgccaggat tacagtgtcc aaagaccaat cctcctgtac
     1921 cacagcccgg acgaagtctc gctctgtcac caggctggag tgcagtggca tgatcttggc
     1981 tcactgcaac ctccgcctcc cgggttcaag agattctcct gcctcagcct cccaagcagc
     2041 tgggactaca gggatgtgcc accaggccga cgccaccagg cccagcgacc cagctttctt
     2101 gtacaaagtg gttgatggta ccgcccgggg acgcacccca cccagcctgg gtccccaaga
     2161 tgagtcctgc accaccgcca gctcctccct ggccaaggac acttcatcgg ccagccctag
     2221 caaccctggc gccagcaacg gcagcggaga gagcttgttt aagggaccac gtgattacaa
     2281 cccgatatcg agcaccattt gtcatttgac gaatgaatct gatgggcaca caacatcgtt
     2341 gtatggtatt ggatttggtc ccttcatcat tacaaacaag cacttgtttc gtcgcaataa
     2401 tggaacactg ttggtccaat cactacatgg tgtattcaag gtcaaggaca ccacgacttt
     2461 gcaacaacac ctcattgatg ggagggacat gatgattatt cgcatgccta aggatttccc
     2521 accatttcct caaaagctga aatttagaga gccacaaagg gaagagcgca tatgtcttgt
     2581 gacaaccaac ttccaagctc ccgagaacct gtacttccag ggttctagag aacagaagct
     2641 gatctcagaa gaggatctgg cccgggccat gaagctactg tcttctatcg aacaagcatg
     2701 cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg
     2761 tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag
     2821 ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat
     2881 ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc
     2941 attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt
     3001 ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc
     3061 atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgc cggaattccc
     3121 ggggatcagc ttgataattc gattcaccat gcccaagaag aagaggaagg tggcgtacag
     3181 ccgcgcgcgt acgaaaaaca attacgggtc taccatcgag ggcctgctcg atctcccgga
     3241 cgacgacgcc cccgaagagg cggggctggc ggctccgcgc ctgtcctttc tccccgcggg
     3301 acacacgcgc agactgtcga cggccccccc gaccgatgtc agcctggggg acgagctcca
     3361 cttagacggc gaggacgtcg cgatggcgca tgccgacgcg ctagacgatt tcgatctgga
     3421 catgttgggg gacggggatt ccccgggtcc gggatttacc ccccacgact ccgcccccta
     3481 cggcgctctg gatatggccg acttcgagtt tgagcagatg tttaccgatg cccttggaat
     3541 tgacgagtac ggtggggcgg ccgccaccat ggggtatcca tatgatgttc cagattatgc
     3601 tagatctgga atcactagtg aattatcgaa ttctgcagcc aagctaattc cgggcgaatg
     3661 tcttatgatt tatgattttt attattaacc tgcagggcgg ccgcgactct agatcataat
     3721 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3781 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3841 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3901 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     3961 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4021 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4081 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4141 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4201 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4261 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4321 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4381 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4441 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4501 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4561 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4621 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4681 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4741 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4801 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4861 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4921 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4981 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5041 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5101 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5161 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5221 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5281 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5341 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5401 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5461 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5521 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5581 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5641 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5701 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5761 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5821 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5881 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5941 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6001 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6061 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6121 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     6181 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6241 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6301 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6361 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6421 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6481 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6541 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6601 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6661 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6721 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6781 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6841 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6901 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6961 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7021 cgccatgcat
//
