LOCUS       sequence-204691-        9016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSIN - EF-1alpha - Cyto -sFd - P2A - Cyto - sFNR, vector encoding 
            for cytoplasmic-tagged Synechococcus sp. Heme Oxigenase-1 (HO1) and 
            Phycocyanobilin:ferredoxin oxireductase (PcyA), under EF-1alpha 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9016)
  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 9016)
  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-240
FEATURES             Location/Qualifiers
     source          1..9016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..1180
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          233..1171
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1128..1148
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1274..1283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1283..1312
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2042..2098
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2102..2125
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2896..2916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2901..3230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3081..3216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3240..3839
                     /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(3240..3259)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3736..3756
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4132..4365
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4516..4533)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4522..4746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4789..4798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4842..4946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4947..5015,5016..5807)
                     /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(5165..5184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5978..6566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6467..6486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             6725..7358
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7405..7530
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8023..8256
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8441..8485
                     /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    8772..8889
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
       61 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      121 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      181 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      241 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      301 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      361 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      421 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      481 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      541 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      601 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      661 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      721 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
      781 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
      841 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
      901 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
      961 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1021 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1081 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1141 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattgatc cttcgaacta
     1201 gaattcgaag cttgagctcg agatctgagt ccggtagcgc tagttcgaag gatccgctag
     1261 cacattcgaa gccgccacca tggaacagaa gctgatctcc gaagaggacc tgatggtcgc
     1321 caacctggcc accatgctga gggagggcac caagacaagt cacaccatgg ccgagaatgt
     1381 cggattcgtg aaatgctttc tcaagggtgt tgtggaaaag aagtcctaca ggaagctggt
     1441 ggctgacctg tattacgtgt actcagccat ggaagaggag atggagagac tgaaagatca
     1501 tcctgtcgtg tcccagatct attttccaga actcaacagg aagcagtcac tggagacaga
     1561 cctgagatat tacttcggac ccaattggca ggccgaggcc aaaattaccc ccgcaggaca
     1621 ggcctatgtg gacaggattc atgaggtggc tcagaccgcc cctgaactcc tggtctcaca
     1681 ctcctatacc cgctatctcg gcgacctgag cggaggccag atcctgaaga agatcgcaca
     1741 gaacgcaatg aatctggacg gggagggaac cgctttctac gagtttgaga atatcagtga
     1801 cgagaaagcc ttcaaagata agtaccgggc agcaatgaat agcctggatg tgccagagga
     1861 aactgctgaa cagatcgtgc aggaagccaa tgatgccttc ggcatgaaca tgaatatgtt
     1921 taaggagctg gaaggtaatc tggtcaaggc catcggcgtc atgctgttta ataccctgac
     1981 acgccggaga accaagggca gcacagacgc agacctgtca cctgcttcta atggaagcgg
     2041 agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctat
     2101 ggattacaag gatgacgacg ataagtctag catgactgct cctgccacaa aacctaagtt
     2161 ttaccctctg attgagcagc tggcaggagt gatcctggag acctggcacc agcatctgga
     2221 gctggaaccc tttaccctgc ctgaaggact gggctatatt gaaggcaagc tggaaggcga
     2281 gagactgact attgagaaca cctgttacca gagccagcac ttcaggaaga tgcacctgga
     2341 gatggccaaa gtgggagaaa acctggacat cctgcactgc gtcatgtttc ccagggccga
     2401 atatgctctg cccatgttcg gctgtgacct ggtcggcaat ccaagaggcg tgagtgccgc
     2461 tattgcagac ctgtctccta cctccgcaga caagactctg tccgtggact atcagacaag
     2521 gctgagacag aaccagggga atctgaactt cagtcagcca agagacctgc ccgaatgggg
     2581 agcaatcttc agcgagtttt gcctgtttat ccggccaagt aatcccaacg aagagcagca
     2641 gtttctggag cgcgtgagag agttcctgac aatccattgt cagctggccc tgggcctgga
     2701 aaagctgagc ccagcagaac aggctatcta tctggctggc cagaggcact attgccagga
     2761 gcagcagagg aatgacaaaa ccaggagagt gctggagaaa gcctttaacc ccgattgggc
     2821 agagcgatac atgagccagg tgctgtttga catccccgcc taataagaat tcgccagcac
     2881 agtggtcgac cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3061 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3121 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttacca
     3241 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     3301 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     3361 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     3421 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     3481 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     3541 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     3601 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     3661 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     3721 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     3781 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     3841 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgataa
     3901 aataaaagat tttatttagt ctccagaaaa agggggcggc cgctcgagac ctagaaaaac
     3961 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
     4021 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
     4081 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     4141 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     4201 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     4261 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     4321 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     4381 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt
     4441 gacattataa tagatttagc aggaattgaa ctaggagtgg agcacacagg caaagttcta
     4501 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4561 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4621 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4681 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4741 ctatggcttc tgaggcggaa agaaccagct gggggcgcgc ccctcgaggc cgccatggtc
     4801 atagctgttt gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4861 tatttttcta aatacattca aatatgtatc cgctcacgag acaataaccc tgataaatgc
     4921 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4981 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5041 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5101 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     5161 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5221 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     5281 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5341 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     5401 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5461 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5521 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5581 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5641 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5701 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5761 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5821 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5881 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6121 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttctggc
     6601 cttttgcacg gaccgccact gccaattacc tgtggtttca tttactctaa acctgtgatt
     6661 cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag
     6721 tagttggaag ggctaattca ctcccaaaga agacaagata tccttgatct gtggatctac
     6781 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat
     6841 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag
     6901 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac
     6961 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc
     7021 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt
     7081 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc
     7141 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat
     7201 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7261 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7321 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7381 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7441 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7501 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7561 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7621 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7681 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7741 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7801 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7861 cagcaaccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7921 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7981 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     8041 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     8101 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     8161 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8221 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8281 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8341 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8401 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8461 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8521 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8581 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8641 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     8701 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccaca aatggcagta
     8761 ttcatccaca attttaaaag aaaggggggg attggggggt acagtgcagg ggaaagaata
     8821 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     8881 caaaattttc gggtttatta cagggacagc agagatccac tttggatcga taagctttgc
     8941 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
     9001 tgaaaggagt gggaat
//
