LOCUS       sequence-166569-        4202 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of photoactivated cyclase bPAC of Beggiatoa sp..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4202)
  AUTHORS   Stierl M, Stumpf P, Udwari D, Gueta R, Hagedorn R, Losi A, Gartner 
            W, Petereit L, Efetova M, Schwarzel M, Oertner TG, Nagel G, Hegemann
            P
  TITLE     Light modulation of cellular cAMP by a small bacterial 
            photoactivated adenylyl cyclase, bPAC, of the soil bacterium 
            Beggiatoa.
  JOURNAL   J Biol Chem. 2011 Jan 14. 286(2):1181-8.
  PUBMED    21030594
REFERENCE   2  (bases 1 to 4202)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2011 Jan 14. 286(2):1181-8."
COMMENT     Sequence Label: bPAC-mycHis_pGEM
FEATURES             Location/Qualifiers
     source          1..4202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14..32
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           64..106
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1203..1232
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1233..1262
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     3'UTR           1311..1436
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1557..1575)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1558..1575)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1593..1609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1593..1609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1606..1628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1617..1633
                     /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(1641..1647,1648..1665,1666..1671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1686..1707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1824..1841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1995..2583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2075..2094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2754..3545,3546..3614))
                     /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     3377..3396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3615..3719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3787..3805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3843..3865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3965..3984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4178..4200
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4192..4202,1..7)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(4193..4202,1..7)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggccagtgaa ttgtaatacg actcactata gggcgaatta attcgagctc ggtacccagc
       61 ttgcttgttc tttttgcaga agctcagaat aaacgctcaa ctttggcaga tcaattcccc
      121 ggggatccaa gcttgccacc atgatgaagc ggctggtgta catcagcaag atcagcggcc
      181 acctgagcct ggaagagatc cagcggatcg gcaaggtgtc catcaagaac aaccagcggg
      241 acaacatcac cggcgtgctg ctgtacctgc agggcctgtt cttccagatc ctggaaggcg
      301 agaacgagaa ggtggacaag ctgtacaaga aaatcctggt ggacgaccgg cacaccaaca
      361 tcctgtgcct gaaaaccgag tacgacatca ccgaccggat gttccccaac tgggccatga
      421 aaaccatcaa cctgaacgag aacagcgagc tgatgatcca gcccatcaag agcctgctgc
      481 agaccatcac ccagagccac cgggtgctgg aaaagtacat gcccgccaga gtgatctacc
      541 tgatcaacca gggcatcaac cccctgaccg tggagcccca gctggtcgag aagatcatct
      601 tcttcagcga catcctggcc ttcagcaccc tgaccgagaa gctgcccgtg aacgaggtgg
      661 tcatcctggt caaccggtac ttcagcatct gcacccggat catcagcgcc tacggcggcg
      721 aagtgaccaa gttcatcggc gactgcgtga tggccagctt caccaaagag cagggcgacg
      781 ccgccatccg gaccagcctg gacatcatct ccgagctgaa gcagctgcgg caccacgtgg
      841 aggccaccaa ccccctgcac ctgctgtaca ccggcatcgg cctgagctac ggccacgtga
      901 tcgagggcaa catgggcagc agcctgaaga tggaccacac cctgctgggg gacgccgtga
      961 acgtggccgc caggctggaa gccctgacaa gacagctgcc ctacgccctg gcctttacag
     1021 ccggcgtgaa gaagtgctgc caggcccagt ggaccttcat caacctggga gcccaccagg
     1081 tcaagggcaa gcaggaagcc atcgaggtct acaccgtgaa tgaggcccag aagtactacg
     1141 acaccctgca gatcacccag ctgatccggc agaccctgga aaacgacaag ccgcggacgt
     1201 acgagcaaaa gctgatctcg gaggaggacc tgcaccatca ccatcaccat caccatcacc
     1261 atctcgagta agcggccgca ggacctgtaa gcttgatctg gttaccacta aaccagcctc
     1321 aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt acaaaatgtt
     1381 gtcccccaaa atgtagccat tcgtatctgc tcctaataaa aagaaagttt cttcacattc
     1441 taaaaaaaaa aaaaaaaaaa aaaacccccc cccccccccc cccccccccc ccccctgcag
     1501 gcatgcgttt aaacatttaa attacgtatt aattaagata tcgctagctt gagtattcta
     1561 tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1621 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1681 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1741 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1801 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1861 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1921 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1981 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2041 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2101 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2161 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2221 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2281 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2341 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2401 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2461 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2521 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2581 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2641 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2701 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2761 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2821 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2881 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2941 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3001 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3061 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3121 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3181 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3241 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3301 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3361 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3421 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3481 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3541 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3601 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3661 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3721 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3781 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     3841 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3901 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     3961 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4021 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     4081 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     4141 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     4201 ac
//
