LOCUS       sequence-403456-        5401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5401)
  AUTHORS   Koch WJ, Hawes BE, Inglese J, Luttrell LM, Lefkowitz RJ
  TITLE     Cellular expression of the carboxyl terminus of a G protein-coupled 
            receptor kinase attenuates G beta gamma-mediated signaling.
  JOURNAL   J Biol Chem. 1994 Feb 25. 269(8):6193-7.
  PUBMED    8119963
REFERENCE   2  (bases 1 to 5401)
  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: "J Biol 
            Chem. 1994 Feb 25. 269(8):6193-7."
COMMENT     Sequence Label: pRK5 BARK1 minigene
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      928..937
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1701..1710
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1714..1848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1751..1770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1805..1824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1917..2274
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1917..2108
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1940..1960)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2125..2260
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2294..2311)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2294..2310)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2303..2325)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2523..2978
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2610..2629)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2820..2841
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2995..3014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3114..3136
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3174..3192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3260..3364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3365..3433,3434..4225)
                     /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     complement(3583..3602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4396..4984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4885..4904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5138..5155
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5272..5293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5308..5313,5314..5331,5332..5338)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5346..5362
                     /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     5351..5373
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5370..5386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5370..5386
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgccgcc accatgggaa tcaagctact ggacagtgac
      961 caggagctct accgcaactt ccccctgacc atctcggagc ggtggcagca ggaggtagca
     1021 gagactgtct ttgacaccat caatgctgag acggaccggc tggaggcccg caagaaaacc
     1081 aaaaacaagc agttgggcca cgaggaagac tacgccctgg gcaaggactg catcatgcat
     1141 ggctacatgt ccaagatggg caaccccttc ctgacccagt ggcagcggcg gtacttctac
     1201 ctgttcccta accggctcga gtggcggggc gagggcgagg ccccgcagag cctgctgacc
     1261 atggaggaga tccagtcggt ggaggagacg cagatcaagg agcgaaagtg cctcctcctc
     1321 aagatccgag gtggcaagca gtttgtcctg cagtgcgata gtgacccaga gctggtgcag
     1381 tggaagaagg agcttcgaga cgcctaccgc gaggcccagc agctagtgca gcgggtgccc
     1441 aagatgaaga acaagccgcg ctcgcccgtc gtggagctga gcaaggtgcc actgatccag
     1501 cgcggcagtg ccaacggcct ctaatgatca gctcgctttc ttgctgtcca atttctatta
     1561 aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga agggccttga
     1621 gcatctggat tctgcctaat aaaaaacatt tattttcatt gcatgatgta tttaatctag
     1681 agtcgacctg cagaagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt
     1741 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1801 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc
     1861 ggcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg taccttctga
     1921 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1981 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2041 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2101 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2161 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2221 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg
     2281 ttaacagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2341 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2401 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     2461 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     2521 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2581 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2641 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2701 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     2761 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2821 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
     2881 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2941 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3001 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3061 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3121 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3181 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3241 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3301 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3361 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3421 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3481 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3541 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3601 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3661 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3721 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3781 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3841 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3901 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3961 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4021 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4081 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4141 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4201 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4261 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4321 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4381 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4441 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4501 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4561 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4621 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4681 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4741 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4801 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4861 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4921 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4981 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5041 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5101 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5161 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     5221 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     5281 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     5341 tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga ttacgaatta
     5401 a
//
