LOCUS       sequence-322368-        7379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for preparation of stable cell lines expressing 
            Barrestin2-TEV fusion protein in lieu of the HTLA cell line..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7379)
  AUTHORS   Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, 
            Sciaky N, Roth BL
  TITLE     PRESTO-Tango as an open-source resource for interrogation of the 
            druggable human GPCRome.
  JOURNAL   Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. 
            Epub 2015 Apr 20.
  PUBMED    25895059
REFERENCE   2  (bases 1 to 7379)
  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"; doi: "10.1038/nsmb.3014";
            journalName: "Nat Struct Mol Biol"; date: "2015-05"; volume: "22"; 
            issue: "5"; pages: "362-9"
COMMENT     Sequence Label: pCDNA3.1(+)-CMV-bArrestin2-TEV
FEATURES             Location/Qualifiers
     source          1..7379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2972..2989)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2978..3202
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3248..3676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3335..3354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3545..3566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3685..3705)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3690..4019
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3870..4005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3932..3951
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4086..4880
                     /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(4140..4159)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4750..4769
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5054..5175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5091..5110)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5145..5164
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5224..5240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5224..5240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5237..5259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5248..5264
                     /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(5272..5278,5279..5296,5297..5302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5317..5338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5455..5472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5626..6214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5706..5725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6385..7176,7177..7245))
                     /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     7008..7027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7246..7350)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgg taccgagctc ggatccacca tgggggagaa acccgggacc
      961 agggtcttca agaagtcgag ccctaactgc aagctcaccg tgtacttggg caagcgggac
     1021 ttcgtagatc acctggacaa agtggaccct gtagatggcg tggtgcttgt ggaccctgac
     1081 tacctgaagg accgcaaagt gtttgtgacc ctcacctgcg ccttccgcta tggccgtgaa
     1141 gacctggatg tgctgggctt gtccttccgc aaagacctgt tcatcgccac ctaccaggcc
     1201 ttccccccgg tgcccaaccc accccggccc cccacccgcc tgcaggaccg gctgctgagg
     1261 aagctgggcc agcatgccca ccccttcttc ttcaccatac cccagaatct tccatgctcc
     1321 gtcacactgc agccaggccc agaggataca ggaaaggcct gcggcgtaga ctttgagatt
     1381 cgagccttct gtgctaaatc actagaagag aaaagccaca aaaggaactc tgtgcggctg
     1441 gtgatccgaa aggtgcagtt cgccccggag aaacccggcc cccagccttc agccgaaacc
     1501 acacgccact tcctcatgtc tgaccggtcc ctgcacctcg aggcttccct ggacaaggag
     1561 ctgtactacc atggggagcc cctcaatgta aatgtccacg tcaccaacaa ctccaccaag
     1621 accgtcaaga agatcaaagt ctctgtgaga cagtacgccg acatctgcct cttcagcacc
     1681 gcccagtaca agtgtcctgt ggctcaactc gaacaagatg accaggtatc tcccagctcc
     1741 acattctgta aggtgtacac cataacccca ctgctcagcg acaaccggga gaagcggggt
     1801 ctcgccctgg atgggaaact caagcacgag gacaccaacc tggcttccag caccatcgtg
     1861 aaggagggtg ccaacaagga ggtgctggga atcctggtgt cctacagggt caaggtgaag
     1921 ctggtggtgt ctcgaggcgg ggatgtctct gtggagctgc cttttgttct tatgcacccc
     1981 aagccccacg accacatccc cctccccaga ccccagtcag ccgctccgga gacagatgtc
     2041 cctgtggaca ccaacctcat tgaatttgat accaactatg ccacagatga tgacattgtg
     2101 tttgaggact ttgcccggct tcggctgaag gggatgaagg atgacgacta tgatgatcaa
     2161 ctctgcggat ccggtgagag tctcttcaaa gggccgcgcg attataaccc catctcatcc
     2221 acaatttgtc acctcacaaa tgagtccgat ggacatacaa caagccttta cggcattggc
     2281 ttcggtccat tcatcatcac caacaagcat cttttcagac gcaataacgg caccttgctg
     2341 gtgcagtccc tccacggcgt ctttaaagtc aagaacacca cgacccttca gcaacacctg
     2401 atcgacggaa gggatatgat tatcattaga atgcccaaag atttcccgcc atttcctcag
     2461 aaactcaagt ttagagagcc acagagggaa gaaaggatct gcttggtgac aacaaatttt
     2521 caaactaagt ctatgtcatc catggtatca gataccagct gtacgtttcc tagttcagac
     2581 ggaatattct ggaagcattg gatacagaca aaggacggcc aatgcggtag tccactggtg
     2641 tctactcggg atggtttcat cgtcggcatt catagcgcca gcaacttcac caatactaat
     2701 aactacttca ccagcgtgcc caagaatttc atggagcttc tgacaaacca ggaagcccag
     2761 cagtgggttt ctggatggcg cttgaacgct gactccgttc tgtggggagg ccataaagtg
     2821 tttatgagta aacctgaaga accctttcag ccggtgaaag aggccactca gcttatgaat
     2881 gagctggttt atagccagta ggaattctgc agatatccag cacagtggcg gccgctcgag
     2941 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3001 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3061 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3121 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3181 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3241 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3301 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3661 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3841 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3901 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3961 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4021 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4081 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4141 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4201 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4261 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4321 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4381 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4441 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4501 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4561 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4621 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4681 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4741 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4801 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4861 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4921 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4981 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5041 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5101 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5161 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5221 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5281 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5341 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5401 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5461 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5521 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5581 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5641 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5701 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5761 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5821 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5881 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5941 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6001 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6061 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6121 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6181 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6241 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6301 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6361 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6421 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6481 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6541 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6601 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6661 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6721 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6781 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6841 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6901 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6961 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7021 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7081 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7141 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7201 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7261 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7321 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
