LOCUS       sequence-492663-        5835 bp    DNA     circular SYN 12-NOV-2025
DEFINITION  Expression of bovine arrestin-3 in bacteria.
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 Nov 12, 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
FEATURES             Location/Qualifiers
     source          1..5835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(296..387)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(406..433)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(525..611)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     764..781
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(2391..2417)
                     /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"
     misc_feature    2468..2537
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(2543..2565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2554..2570
                     /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(2578..2584,2585..2601,2602..2607))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(2839..3921)
                     /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     3883..3902
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3922..3999)
                     /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."
     primer_bind     complement(4077..4099)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4185..4325
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4240..4259
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4340..4357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4511..5099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4591..4610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5270..5835,1..226,227..295))
                     /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"
ORIGIN
        1 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
       61 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
      121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
      181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
      241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      361 ttgaatgtat ttagaaaaat aaacaaaaga gtttgtagaa acgcaaaaag gccatccgtc
      421 aggatggcct tctgcttaat ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
      481 cctccgggcc gttgcttcgc aacgttcaaa tccgctcccg gcggatttgt cctactcagg
      541 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
      601 cgttttattt gatgcctggc agttccctac tctcgcatgg ggagacccca cactaccatc
      661 ggcgctacgg cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact
      721 gccgccaggc aaattctgtt ttatcagacc gcttctgcgt tctgatttaa tctgtatcag
      781 gctgaaaatc ttctctcatc cgccaaaaca gccaagcttg atatcgaatt cggctttccc
      841 cctcttttcc tactatcact cctcttcggc tccccctcca aaaaattgag gtgtgaacac
      901 agcttgccac ccactggggg attggggaag cacagctaaa gtcaaaaaat taatgcccat
      961 gttccacaag agctgggtcc agtgcgtatc tggtgggggg gtagggcagg gaggggggga
     1021 ggggttggtc gaggaagctg gtgggtggag gggaagggaa gagggactgg gatccccctc
     1081 agtgacagtg ggggtcttgg tcctgcctcc acctgtcccc cgccccgcca tccactccct
     1141 tcttccagcc tcccttccta gcagaactgg tcgtcatagt cctcgtcctt cagccccttg
     1201 agccgcagcc gggcaaagtc ctcaaacacg atgtcgtcgt ctgtggcata gttggtttcg
     1261 aattcaatga ggttggtgtc cacgggggca tctgtttcag gaacagctga ctggggcctg
     1321 ggaagggcga tgtgatcatg gggcttgggg tgcatgagaa caaatggcag ctccactgag
     1381 acgtccccgc ctcgagacac caccagcttc accttgaccc tgtaggacac caggatcccc
     1441 agcacctcct tgttggcacc ctccttcaca atggtgctgg acgccaggtt ggtgtcctcg
     1501 tgcttgagct tcccatccag agcgaggccg cgcttctccc ggttgttgct gagcagcggg
     1561 gttatggtgt acaccttaca gaacgtggaa ctgggcgaca cctggtcatc ttgttcgacc
     1621 tgagccacgg ggcacttgta ctgggcggtg ctgaagaggc agatgtcggc gtactgtctc
     1681 acagagactt tgatcttctt gacagtcttg gtggagttat tggtgacgtg gacattgaca
     1741 ttgaggggct ccccatggta gtacagctcc ttgtccagtg aagcctcaag gtgcagggac
     1801 cggtcagaca tgaggaagtg gcgtgtggtt tcagctgaag gctgggggcc gggtttctct
     1861 ggggcaaact gcacctttcg gatcaccaga cgcacagagt tccgtttgtg gcttttctct
     1921 tctagtgatt tggcgcagaa ggctcgaatc tcaaagtcta ccccgcaggc cttccccgta
     1981 tcctctgggc ctggctgcag ggtgacggag cagggcaggt tctggggtat tgtgaaaaaa
     2041 aaggggtggg catgctggcc cagcttcctc agcagccgct cctgcaggcg ggtggggggc
     2101 cggggtgggt tgggtgttgg ggggaaggcc tggtagttgg cgatgaacag gtctttgcgg
     2161 aaggacaagc ccagcacgtc caggtcttcg cggccatagc gaaaggcgca ggtgagggtc
     2221 acaaacactt tgcggtcctt caagtagtcc gggtccacta gcaccacacc atctacagga
     2281 tccactttgt ccaggtggtc cacaaagtcc cgcttgccca ggtacacggt gagcttgcag
     2341 ttaggactcg atttcttgaa gaccctggtc ccgggtttct cccccatggt ttattcctcc
     2401 ttatttaatc gatacattaa tatatacctc tttaattttt aataataaag ttaatcgata
     2461 attccggtcg agtgcccaca cagattgtct gataaattgt taaagagcag tgccgcttcg
     2521 ctttttctca gcggcgctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacat
     2581 tatacgagcc ggatgattaa ttgtcaacag ctcatttcag aatatttgcc agaaccgtta
     2641 tgatgtcggc gcaaaaaaca ttatccagaa cgggagtgcg ccttgagcga cacgaattat
     2701 gcagtgattt acgacctgca cagccatacc acagcttccg atggctgcct gacgccagaa
     2761 gcattggtgc accgtgcagt cgatgataag ctgtcaaacc agatcaattc gcgctaactt
     2821 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2881 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     2941 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     3001 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     3061 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     3121 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     3181 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     3241 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     3301 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     3361 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     3421 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     3481 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     3541 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     3601 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     3661 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     3721 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     3781 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     3841 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     3901 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     3961 tgccataccg cgaaaggttt tgcaccattc gatggtgtca acgtaaatgc atgccgcttc
     4021 gccttcgcgc gcgaattgat ctgctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4081 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4141 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4201 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4261 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4321 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4381 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4441 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4501 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4561 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4621 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4681 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4741 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4801 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4861 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4921 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     4981 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5041 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5101 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5161 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5221 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5281 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5341 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5401 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5461 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5521 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5581 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5641 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5701 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5761 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5821 agtactcaac caagt
//
