LOCUS       sequence-471663-        5835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of bovine arrestin-3 short (1-392) L393Stop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5835)
  AUTHORS   Vishnivetskiy SA, Zhan X, Gurevich VV
  TITLE     Expression of Untagged Arrestins in E. coli and Their Purification.
  JOURNAL   Curr Protoc. 2023 Sep;3(9):e832. doi: 10.1002/cpz1.832.
  PUBMED    37671938
REFERENCE   2  (bases 1 to 5835)
  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: "Curr 
            Protoc."; date: "2023-09"; volume: "3(9):e832. doi"; pages: " 
            10.1002/cpz1.832"
COMMENT     Sequence Label: BtARR3 short (1-392)
FEATURES             Location/Qualifiers
     source          1..5835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    263..332
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             383..409
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     primer_bind     complement(2019..2036)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2189..2275
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2367..2394
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2413..2504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2505..2573,2574..3365)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2723..2742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3536..4124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4025..4044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4278..4295
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4310..4450
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4376..4395)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4536..4558
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4636..4713
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4714..5796
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4733..4752)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatggggga
      421 gaaacccggg accagggtct tcaagaaatc gagtcctaac tgcaagctca ccgtgtacct
      481 gggcaagcgg gactttgtgg accacctgga caaagtggat cctgtagatg gtgtggtgct
      541 agtggacccg gactacttga aggaccgcaa agtgtttgtg accctcacct gcgcctttcg
      601 ctatggccgc gaagacctgg acgtgctggg cttgtccttc cgcaaagacc tgttcatcgc
      661 caactaccag gccttccccc caacacccaa cccaccccgg ccccccaccc gcctgcagga
      721 gcggctgctg aggaagctgg gccagcatgc ccaccccttt tttttcacaa taccccagaa
      781 cctgccctgc tccgtcaccc tgcagccagg cccagaggat acggggaagg cctgcggggt
      841 agactttgag attcgagcct tctgcgccaa atcactagaa gagaaaagcc acaaacggaa
      901 ctctgtgcgt ctggtgatcc gaaaggtgca gtttgcccca gagaaacccg gcccccagcc
      961 ttcagctgaa accacacgcc acttcctcat gtctgaccgg tccctgcacc ttgaggcttc
     1021 actggacaag gagctgtact accatgggga gcccctcaat gtcaatgtcc acgtcaccaa
     1081 taactccacc aagactgtca agaagatcaa agtctctgtg agacagtacg ccgacatctg
     1141 cctcttcagc accgcccagt acaagtgccc cgtggctcag gtcgaacaag atgaccaggt
     1201 gtcgcccagt tccacgttct gtaaggtgta caccataacc ccgctgctca gcaacaaccg
     1261 ggagaagcgc ggcctcgctc tggatgggaa gctcaagcac gaggacacca acctggcgtc
     1321 cagcaccatt gtgaaggagg gtgccaacaa ggaggtgctg gggatcctgg tgtcctacag
     1381 ggtcaaggtg aagctggtgg tgtctcgagg cggggacgtc tcagtggagc tgccatttgt
     1441 tctcatgcac cccaagcccc atgatcacat cgcccttccc aggccccagt cagctgttcc
     1501 tgaaacagat gcccccgtgg acaccaacct cattgaattc gaaaccaact atgccacaga
     1561 cgacgacatc gtgtttgagg actttgccta gctgcggctc aaggggctga aggacgagga
     1621 ctatgacgac cagttctgct aggaagggag gctggaagaa gggagtggat ggcggggcgg
     1681 gggacaggtg gaggcaggac caagaccccc actgtcactg agggggatcc cagtccctct
     1741 tcccttcccc tccacccacc agcttcctcg accaacccct ccccccctcc ctgccctacc
     1801 cccccaccag atacgcactg gacccagctc ttgtggaaca tgggcattaa ttttttgact
     1861 ttagctgtgc ttccccaatc ccccagtggg tggcaagctg tgttcacacc tcaatttttt
     1921 ggagggggag ccgaagagga gtgatagtag gaaaagaggg ggaaagccga attcgatatc
     1981 aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa
     2041 cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct
     2101 gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc
     2161 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2221 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     2281 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     2341 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
     2401 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     2761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3661 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4321 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     4381 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4441 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4501 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4561 ttcaccgtca tcaccgaaac gcgcgaggca gcagatcaat tcgcgcgcga aggcgaagcg
     4621 gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag
     4681 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     4741 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     4801 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     4861 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     4921 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     4981 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     5041 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     5101 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     5161 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     5221 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     5281 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     5341 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     5401 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     5461 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     5521 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     5581 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     5641 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     5701 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     5761 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     5821 gcgcgaattg atctg
//
