LOCUS       sequence-204688-        7849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pSIN - EF-1alpha - MTS - tFd, vector encoding 
            mitochondrial-tagged Thermosynechococcus elongates Ferredoxin (Fd), 
            under EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7849)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, 
            Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and 
            Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded 
            PhyB Optogenetics.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 
            10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
  PUBMED    29301067
REFERENCE   2  (bases 1 to 7849)
  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"; doi: 
            "10.1021/acssynbio.7b00413"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "706-717"
COMMENT     Sequence Label: pPKm-300
FEATURES             Location/Qualifiers
     source          1..7849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1264..1338
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      1360..1369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1369..1410
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1728..1748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1733..2062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1913..2048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2072..2671
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2072..2091)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2568..2588
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2964..3197
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(3348..3365)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3354..3578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      3621..3630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3779..3847,3848..4639)
                     /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(3997..4016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5557..6190
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6237..6362
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6855..7088
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7273..7317
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7604..7721
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattgatcc ttcgaactag
     1201 aattcgaagc ttgagctcga gatctgagtc cggtagcgct agttcgaagg atccgccgcc
     1261 accatgagcg tgctgacgcc cttgctgctt agaggactga ccggaagcgc acgccgcctg
     1321 cccgtgccta gggcgaagat acatagcctg ttcgaagccg ccaccatggg taagcccatt
     1381 cccaacccgc tgttgggcct cgatagcaca gttaacatgg ctacctacaa agtgacactg
     1441 gttcggcctg atggtagcga gacaacaatt gacgtgcctg aggacgaata catactcgac
     1501 gttgcggaag agcaagggct ggatctgcca tttagttgcc gagcgggcgc ttgcagcact
     1561 tgtgccggga agttgctcga aggggaagtg gatcagtctg atcagtcctt tctggatgac
     1621 gatcagatcg aaaaagggtt cgtgctcacc tgtgtcgctt atccaaggag cgattgtaag
     1681 attctcacta atcaggagga ggagctttac taataagtcg accctgtgga atgtgtgtca
     1741 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1801 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1861 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     1921 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     1981 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2041 tggaggccta ggcttttgca aaaagcttac catgaccgag tacaagccca cggtgcgcct
     2101 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     2161 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     2221 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     2281 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     2341 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     2401 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     2461 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     2521 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     2581 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     2641 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     2701 cgaaaggagc gcacgacccc atgcatcgat aaaataaaag attttattta gtctccagaa
     2761 aaagggggcg gccgctcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     2821 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     2881 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     2941 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     3001 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3061 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3121 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     3181 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     3241 aaagaaatga atatcagaga gtgagaggcc ttgacattat aatagattta gcaggaattg
     3301 aactaggagt ggagcacaca ggcaaagttc tagagctcgc tgatcagcct cgactgtgcc
     3361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3541 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3601 ctgggggcgc gcccctcgag gccgccatgg tcatagctgt ttgacgtcag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcacg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cgccagcaac gcggcctttt tacggttctg gccttttgca cggaccgcca ctgccaatta
     5461 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     5521 ttgtatttgt taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa
     5581 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc
     5641 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc
     5701 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt
     5761 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt
     5821 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga
     5881 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg
     5941 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt
     6001 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6061 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6121 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6181 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     6241 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     6301 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     6361 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     6421 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     6481 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     6541 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     6601 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     6661 aggaagagca aaacaaaagt aagaccaccg cacagcaacc ggccgctgat cttcagacct
     6721 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     6781 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     6841 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     6901 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     6961 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     7021 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     7081 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     7141 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     7201 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     7261 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     7321 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     7381 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     7441 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     7501 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     7561 ggatctcgac ggtatcgcca caaatggcag tattcatcca caattttaaa agaaaggggg
     7621 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     7681 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     7741 gcagagatcc actttggatc gataagcttt gcaaagatgg ataaagtttt aaacagagag
     7801 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaatt
//
