LOCUS       sequence-377048-        6494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aadA.co-TaadA-PJ23117-eYFP.co between homology arms for pilA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6494)
  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 6494)
  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: pJMH143
FEATURES             Location/Qualifiers
     source          1..6494
                     /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"
     promoter        2452..2486
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     protein_bind    2489..2505
                     /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             2523..2534
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(4273..4289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4273..4289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4297..4313
                     /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(4321..4327,4328..4345,4346..4351))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4366..4387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4504..4521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4675..5263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4755..4774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5434..6225,6226..6294))
                     /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     6057..6076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6295..6399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6467..6485
                     /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 gatccgggga gttgacagct
     2461 agctcagtcc tagggattgt gctagctatt gtgagcggat aacaatttca cacatactag
     2521 agaaagagga gaaatactaa tggtatctaa aggtgaagag ctgtttaccg gagttgtacc
     2581 cattttagtt gaattggatg gcgacgtaaa cgggcataaa ttctcagtct ccggggaagg
     2641 agagggagat gcaacttatg gcaaactaac actcaaattt atctgtacca ctgggaaact
     2701 accagtcccc tggcctactc tcgtgacaac ctttggctat gggttacaat gcttcgcaag
     2761 gtatccagat cacatgaaac agcacgactt tttcaaatca gcgatgcctg aaggctacgt
     2821 tcaagagcga acaatatttt tcaaagatga cggaaactac aaaacaagag cggaagttag
     2881 gtttgagggt gataccctgg taaaccgcat cgaattgaaa ggaatcgatt tcaaagagga
     2941 cggtaacata cttggacata aactggaata caactacaac tcgcacaacg tctacatcat
     3001 ggctgataaa cagaaaaatg gtatcaaagt gaacttcaaa atcagacata acatcgagga
     3061 tgggtcagtt cagttagccg accactacca acagaatact ccgatcggtg atggaccggt
     3121 tttattgcca gacaaccatt atctttctta ccagagtgct ctgagcaaag atcctaatga
     3181 aaaacgggac cacatggtcc ttctggagtt tgtgaccgct gccggcatta ctctagggat
     3241 ggatgaactc tataaataac agataatcta gcagtttagg tttcagatgg agtcagctag
     3301 ggtctttaac cccatcgtca actataagat ggagagatgg catacggtga cactgttaaa
     3361 gccataacta tattgctctt gttgataaaa tcagaaactg aggccacttg agagtattgt
     3421 aaataggtag tacagtagtg ctacctattt tgttttaaaa taactaattg agtttatcgc
     3481 tgggactatg gttgataatc agatgtcgga aaagcagaat attttatcag cagaagctga
     3541 tatttacttg cataaaaggg attatcaaca agcagccatt ttatatgaac aagccatcaa
     3601 tctcaatccc gactataaat ataattactg gaaactgggg ttaaccctgc tgctacagga
     3661 gcgggaggaa gaagcccaga ccacctggct ggtggcgatg agtgatggag acccagaaca
     3721 gattgaccag tggacgatag acttactcga ggtgctatca tcagaagccg atgcacgaac
     3781 ctccgccgaa gactattccg tggcttgggg gattcgccaa cacattttag aaataaaccc
     3841 ccatggtatt gataatatat taaagttaat tgatttgtca attaccctga caacctatac
     3901 cggaaatgag atcgcagaat atggactggt tcccttactg caacaggagt ccatcaatgt
     3961 caatccagat ttactactgc acctgtggaa aaaactccta gttgtagatg caattaatcc
     4021 cctatctctg gaattgggag aggcggtgat taaccatctt ggtgagcaac cggagtttgt
     4081 cactagctta atccctaaat tatacgatat tgcctattcc tttggtgaaa taggtatagc
     4141 caagggctat ggagagttgt tgttgaacct ggcaccgcat catcaagagt tgctgagggc
     4201 tctggctcaa ttttgtattc aattaaccga gtatgaccat ggcattgaat atgccagact
     4261 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4321 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4381 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4441 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5101 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5341 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5401 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5461 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5521 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5581 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5641 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5701 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5761 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5821 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5881 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5941 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6001 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6061 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6121 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6181 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6241 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6301 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6361 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6421 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6481 gaggcccttt cgtc
//
