LOCUS       sequence-377053-        6902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aadA.co-TaadA-PcpcB-eYFP.co between homology arms for pilA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6902)
  AUTHORS   Jacob M. Hilzinger, Skyler Friedline, Divya Sivanandan, Ya-Fang 
            Cheng, Shunsuke Yamazaki, Douglas S. Clark, Jeffrey M. Skerker, Adam
            P. Arkin
  TITLE     Acetaminophen production in the edible, filamentous cyanobacterium 
            Arthrospira platensis
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 6902)
  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: "bioRxiv"
COMMENT     Sequence Label: pJMH058
FEATURES             Location/Qualifiers
     source          1..6902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4681..4697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4681..4697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4694..4716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4705..4721
                     /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(4729..4735,4736..4753,4754..4759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4774..4795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4912..4929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5083..5671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5163..5182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5842..6633,6634..6702))
                     /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     6465..6484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6703..6807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6875..6893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtggaat cttgtgcgaa ttgaggtaac
      421 ctcagatcct atccttgagg ttgattgcgt atttctgggg aaaacggagt ttcctcggcc
      481 ctataatgaa actgataagg aggaagattg atgcgaaaat acatcatttt tcgggctgat
      541 aagggtgaac ctggttggaa agaacgtaaa ctccaacata cccagggttt aactagaatt
      601 ttggctgaac atttcgactc ttctgatcaa cctattcctg agttgggata tcgaccgagg
      661 gagtttattc gtattgatgg gttgcataat cccactgaac atggctacag cacccattat
      721 cgtcagggtg actgggaagt aactagggta gaaagttact ctcctgatgt gtctatggct
      781 gagtttgata tgatcgtgat ttgcttttgc aaatatagcc ctgttaatgg tcctgtgcaa
      841 cctatgccag agcgtcgagt gtctgtagat gggttcgctg gggatgagaa gcggttggag
      901 caatggcacg agtcccaaaa gaaaccgact gaggtttagt tggtccccgg tttgcttatt
      961 tgactagcca gattcgccac gacaacagaa ccgccagcag ggaagagggg aatagaaccg
     1021 cgaaaagagc gatcgcattg gtttcgggaa tggcgaaatt tggcccgcca tatttgatca
     1081 gtaaggatag ccctagggat aggatcagga ttttggccac caataacagt ttgttgtcca
     1141 tttatatata ggatgcgatg ggatgcgatc gcctatattt taccaattgg tggcggcctt
     1201 aaccccctaa gccctatgtc tgacacccag tcaccccaac cgccttagtc ggtattcccc
     1261 actgggtggg cgatacacga tttcccggaa aaattgaaaa aaaattttgt ggcaggtgat
     1321 atgatttgcg atcgaggtgg gtaagttaga actacagaaa acaaataacc tcatttcacc
     1381 ccgaggagaa cgaattatga gagaagctgt gatcgccgag gtttcaacac agttatccga
     1441 agttgtaggg gttatcgaac gacatttaga gccaacctta ttggcggtac acttgtatgg
     1501 ttcagctgtc gatggaggcc ttaaacccca ttccgatatt gaccttctgg tcactgtgac
     1561 agttagactt gatgaaacca ctagaagggc cctgatcaat gacttattag aaacttctgc
     1621 aagtcctggt gaaagtgaga tattgagggc ggtagaagtc acaattgtcg tgcacgatga
     1681 cattatcccc tggagatacc ccgctaaaag ggaactgcaa tttggagagt ggcagcgaaa
     1741 cgatattcta gcaggcatat tcgaacctgc gactatcgat attgaccttg ctatactgtt
     1801 gacaaaagcc cgcgagcatt ctgtggcact agttggaccg gctgccgaag aattatttga
     1861 tcctgttccg gaacaagacc ttttcgaagc actgaatgag accctaactc tctggaacag
     1921 tccacccgat tgggccggag acgagcggaa tgttgtatta accttgagcc gaatctggta
     1981 ttcggcggtg actggcaaaa tagctccaaa agatgttgca gcggactggg ctatggaaag
     2041 gttacctgca caatatcagc cggttatact agaggcacgg caagcgtacc tcgggcagga
     2101 agaggatcgc ttagcttcaa gagccgacca attggaagag ttcgtacact acgtcaaagg
     2161 ggaaattacc aaagtcgtgg gtaaataatg tctaacaatt cgttcaagcc gacgccgctt
     2221 cgcggcgcgg cttaactcaa gcgttagatg cactaagcac ataattgctc acagccaaac
     2281 tatcaggtca agtctgcttt tattattttt aagcgtgcat aataagccct acacaaattg
     2341 ggagatatat catgaaaggc tggctttttc ttgttatcgc aatagttggc gaagtaatcg
     2401 caacatccgc attaaaatct agcgagggct ttactaagct gatccggctg cgggattatc
     2461 ccacggttca aggggtggtg gtatttttgg ccgttattgt ggtggcggct agtttggcga
     2521 tcgatatttt gaatgcttgt attgacccac ggattcgcta ttaattccct gtcagaaaat
     2581 atacttagtt attaatactt agttattttt cttaataaat tttaacaaac caaagggcgt
     2641 tggcagtgta taaaggatat ttgaaggggt gagaaggaag gcgaatgttg atttatgaag
     2701 tttgattaac atttgtatca aaatataaaa ttcttctcat aaaccctgta caatctttta
     2761 agatttcgga aagtgttcta ggatactgaa gaaatgaacc acggggcaat tgttaaaagc
     2821 ctttgtcgat ggttcgcccc ggaaggggtc ttaggaggtg acaccgatgg attgattgtc
     2881 gtgatcattc atggtgtgtc caatcccaac tcaactctaa gcaagtcaac aagtaggaga
     2941 taaatccatg gtatctaaag gtgaagagct gtttaccgga gttgtaccca ttttagttga
     3001 attggatggc gacgtaaacg ggcataaatt ctcagtctcc ggggaaggag agggagatgc
     3061 aacttatggc aaactaacac tcaaatttat ctgtaccact gggaaactac cagtcccctg
     3121 gcctactctc gtgacaacct ttggctatgg gttacaatgc ttcgcaaggt atccagatca
     3181 catgaaacag cacgactttt tcaaatcagc gatgcctgaa ggctacgttc aagagcgaac
     3241 aatatttttc aaagatgacg gaaactacaa aacaagagcg gaagttaggt ttgagggtga
     3301 taccctggta aaccgcatcg aattgaaagg aatcgatttc aaagaggacg gtaacatact
     3361 tggacataaa ctggaataca actacaactc gcacaacgtc tacatcatgg ctgataaaca
     3421 gaaaaatggt atcaaagtga acttcaaaat cagacataac atcgaggatg ggtcagttca
     3481 gttagccgac cactaccaac agaatactcc gatcggtgat ggaccggttt tattgccaga
     3541 caaccattat ctttcttacc agagtgctct gagcaaagat cctaatgaaa aacgggacca
     3601 catggtcctt ctggagtttg tgaccgctgc cggcattact ctagggatgg atgaactcta
     3661 taaataacag ataatctagc agtttaggtt tcagatggag tcagctaggg tctttaaccc
     3721 catcgtcaac tataagatgg agagatggca tacggtgaca ctgttaaagc cataactata
     3781 ttgctcttgt tgataaaatc agaaactgag gccacttgag agtattgtaa ataggtagta
     3841 cagtagtgct acctattttg ttttaaaata actaattgag tttatcgctg ggactatggt
     3901 tgataatcag atgtcggaaa agcagaatat tttatcagca gaagctgata tttacttgca
     3961 taaaagggat tatcaacaag cagccatttt atatgaacaa gccatcaatc tcaatcccga
     4021 ctataaatat aattactgga aactggggtt aaccctgctg ctacaggagc gggaggaaga
     4081 agcccagacc acctggctgg tggcgatgag tgatggagac ccagaacaga ttgaccagtg
     4141 gacgatagac ttactcgagg tgctatcatc agaagccgat gcacgaacct ccgccgaaga
     4201 ctattccgtg gcttggggga ttcgccaaca cattttagaa ataaaccccc atggtattga
     4261 taatatatta aagttaattg atttgtcaat taccctgaca acctataccg gaaatgagat
     4321 cgcagaatat ggactggttc ccttactgca acaggagtcc atcaatgtca atccagattt
     4381 actactgcac ctgtggaaaa aactcctagt tgtagatgca attaatcccc tatctctgga
     4441 attgggagag gcggtgatta accatcttgg tgagcaaccg gagtttgtca ctagcttaat
     4501 ccctaaatta tacgatattg cctattcctt tggtgaaata ggtatagcca agggctatgg
     4561 agagttgttg ttgaacctgg caccgcatca tcaagagttg ctgagggctc tggctcaatt
     4621 ttgtattcaa ttaaccgagt atgaccatgg cattgaatat gccagactgg cgtaatcatg
     4681 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4741 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4801 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4861 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5521 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5701 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5761 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5821 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5881 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5941 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6001 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6061 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6841 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6901 tc
//
