LOCUS       sequence-314113-        7586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Heme Oxygenase (Nostocaceae) and ReBphP-PCM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7586)
  AUTHORS   Mishra K, Stankevych M, Fuenzalida-Werner JP, Grassmann S, Gujrati 
            V, Huang Y, Klemm U, Buchholz VR, Ntziachristos V, Stiel AC
  TITLE     Multiplexed whole-animal imaging with reversibly switchable 
            optoacoustic proteins.
  JOURNAL   Sci Adv. 2020 Jun 12;6(24):eaaz6293. doi: 10.1126/sciadv.aaz6293. 
            eCollection 2020 Jun.
  PUBMED    32582850
REFERENCE   2  (bases 1 to 7586)
  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: "Sci Adv."; 
            date: "2020-06-12"; volume: "6(24):eaaz6293. doi"; pages: " 
            10.1126/sciadv.aaz6293. eCollection 2020 Jun"
COMMENT     Sequence Label: pETduet-1:MCS1(HemeOxygenase)::MCS2(ReBphP-PCM)
FEATURES             Location/Qualifiers
     source          1..7586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        240..258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    259..283
                     /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)."
     RBS             298..320
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        1133..1151
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1152..1176
                     /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)."
     CDS             2803..2820
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2873..2891)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2887..2934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2971..3426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3058..3077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3268..3289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3541..4332,4333..4401))
                     /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     4164..4183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4402..4494)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4575..5163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5064..5083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5317..5334
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5349..5488
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5414..5433)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5574..5596
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5590..5781)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6353..7435)
                     /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     7397..7416
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7436..7513)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
       61 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
      121 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      181 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt
      241 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      301 gtttaacttt aagaaggaga tataccatgg cgatgagcag caatttagca aacaaactac
      361 gtgtaggtac gaaaaaagcc cacacaatgg cagaaaatgt aggttttgtc aaatgcttct
      421 taaaaggagt ggtagaaaaa agctcttacc ggaagctagt ggccaacttc tactacgtct
      481 actccgcaat ggaagaggag atggaaaagc atagccagca cccaattgtt tctaaaataa
      541 acttttccca actgaatcgt aagcagactc tagagcaaga cctgagttac tattacggcg
      601 ctaactggcg ggaacaaatt caactgtcgc cagcaggtga agcttacgta caacgcattc
      661 gggaaatctc tgcaaccgaa ccagaattgt tgattgccca ctcttacacc cgttatttag
      721 gtgatttatc cgggggacaa attctcaaaa acattgctgt gacagcgatg aatttgaatg
      781 atgggcaagg aactgcattt tatgagttcg cagatatttc tgatgagaaa gcttttaagg
      841 ctaaatatcg tcaaacatta gatgagttgg caattgatga agcgacaggc gatcgcattg
      901 ttgatgaagc taacgccgct tttggtatga acatgaaaat gttccaagaa ctagaaggga
      961 acctcatcaa ggcaatcggc atgatgctgt tcaacacctt gactcgcaag cgcacacgcg
     1021 gcgctaccga acttgccacg gctgagtaac tgcaggtcga caagcttgcg gccgcataat
     1081 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
     1141 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
     1201 taagaaggag atatacataa acatatgagc ggcaccagag aacccatgga cctgaccaac
     1261 tgcgacagag agcctatcca tcagctgggc gctatccagc ctttcggctt cctgctgcaa
     1321 gtgtccagcg actggatcgt gacaagagcc tccgtgaacc tggccgagtt tctgggagtg
     1381 acacaggccg atgctctggg cagacctgcc agcacactga tcatgcctga ggctctgcac
     1441 accatccgga acaagctgac cacactgaga ggcgccgacg tggtggaaag agtgttcgcc
     1501 attgctctga cccctgacca gagcaagttc gacgtggccg tgcacttcaa cgagagccaa
     1561 gtgatcatcg agggcgagag atgccaagag gacagaagaa acgcccctag cctgagcatg
     1621 cggagcatga tgagcagact ggaccaggcc gagacacttg aggccttttt cagagagggc
     1681 gccagacagg ctagagccct gaccggattc gacagagtga tggtgtacag attcgacgaa
     1741 ggcggctctg gcgaagtggt ggctgaagct gctagaagcg gcatcggcag ctttctgggc
     1801 ctgcactatc ctgccagcga catccctgtg caggccagag cactgtacct gcggaacctg
     1861 ttcagaatca ttgccgatgt ggacgccgtg cctgtgccaa ttctgcccga gagagatgag
     1921 cacggccagc ctctggatct gagcatgagc gtgctgagaa gcgtgtcccc tatccacatc
     1981 gagtacctga agaacatggg cgtgggcgcc agcctgtcta tcagcattgt ggtggatggc
     2041 aagctgtggg gcctgttcgc ctgtcaccac tacgaaccta gactgcccag cgctcagagc
     2101 agatctaccg ccgaactgtt tggccagatg ttcgccagca ggctggaaag cagagagaga
     2161 aggctggccc tggactacga gacaaaggcc agaagaatcg ccgaccggct gctgacatcc
     2221 gtggccgata atgcctctct gctggacgat cctgcctggc tgattgaagc cctggccgat
     2281 gccattcctg ccgatggaat cggcgtgtgg atcaacggta gactggctct ggccggcatc
     2341 ggacccgaca agaagaattt tgccagcctc gtgcggcacc tgaacagaaa tgctggcggc
     2401 agaatctacg ccgtggacag actgagccag acataccccg acctggaaat cgatgctgtg
     2461 gtggccggaa tgctggctat ccccatcagc agatccccac gggattacgt ggtgctgttc
     2521 cggcaagaac tcgtgcggac agttagatgg ggcggagatc ctcacaagcc cgtggaatac
     2581 ggccctaatg gccctagact gacccctcgg aagtccttcg aagagtggag cgaacttgtg
     2641 cggggcagaa gcctgccttt taccgaggct gaacagagag tggccgaaac catcagagtg
     2701 accctgatcg aggtggtgct gagactgaca gacgaggcct ctatggccag acagatggcc
     2761 aatgagagac aagagctgct gatcgccgag ctgaacctcg agcaccacca ccaccaccac
     2821 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     2881 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     2941 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3001 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3061 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3121 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3181 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3241 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3301 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3361 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3421 caatttctgg cggcacgatg gcatgagatt atcaaaaagg atcttcacct agatcctttt
     3481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4381 acggaaatgt tgaatactca tactcttcct ttttcaatca tgattgaagc atttatcagg
     4441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggtc
     4501 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4561 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4621 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4681 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4741 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4801 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4861 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4921 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4981 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5041 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5101 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5161 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5221 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5281 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5341 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     5401 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     5461 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     5521 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5581 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5641 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5701 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5761 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5821 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5881 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     5941 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     6001 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     6061 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     6121 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     6181 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgct
     6241 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     6301 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     6361 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6421 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     6481 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     6541 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     6601 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     6661 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     6721 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     6781 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     6841 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     6901 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     6961 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     7021 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     7081 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     7141 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     7201 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     7261 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     7321 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     7381 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     7441 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     7501 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     7561 cccagtagta ggttgaggcc gttgag
//
