LOCUS       sequence-377047-        8590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aadA.co-TaadA-J23107-4ABH.co.rm1-PpsaA-nhoA.co.rm1 between homology 
            arms for pilA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8590)
  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 8590)
  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: pJMH147
FEATURES             Location/Qualifiers
     source          1..8590
                     /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_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     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(6369..6385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6369..6385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    6393..6409
                     /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(6417..6423,6424..6441,6442..6447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6462..6483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6600..6617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6771..7359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6851..6870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7530..8321,8322..8390))
                     /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     8153..8172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8391..8495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8563..8581
                     /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 gtttacggct
     2461 agctcagccc taggtattat gctagctatt gtgagcggat aacaatttca cacatactag
     2521 agaaagagga gaaatactaa tgtctcaaca agaaagaact agggttgcaa ttgttggagc
     2581 ggggattgtt gggttaacat tggcaattgc gttgaatgcg tttgataaag aaagaaaatt
     2641 ggctatcgat atctatgaaa atgctagtga attggccgaa attggtgcag gaattaatgt
     2701 ttggccaagg actttagcga tcttcaaaca aatcggtgtt gaagatgctt tgattccctt
     2761 attggatcat attcctgatt tagaaccacg gattattttt gggattagaa aaggtgatga
     2821 gaaaaatgga tatcaagttt acgatacaat gaataatggg ggggctttaa gagttcatcg
     2881 agcacatttg caaaacacct tgatccaaca tttaccattg cccgggagca aagttactga
     2941 aatcaactca atctgtggat tccatttggg gcataacttg atcgattact cacatcattc
     3001 atcctctggg caaggtccat taaccttgca tttttcagat ggtaaaccct cccggacttg
     3061 tgatattttg gttggggctg atggaatcaa atccacctta agacatttat ttttgccaag
     3121 gttgcctaac ccagaaaaat acttgaactg ctacgaaccc aaatggaaag ggttattggc
     3181 atatagaggt ttggttccta aagaaaaatt agaagccgtt agtcccggac atagggcatt
     3241 gactcatcct ggtttaatgt atagtggaaa aagcgcttat gccgttgttt accctgttag
     3301 caatgggaaa ttcatcaacg ttgttgccat cgttcatgat aatccaacca attccactgt
     3361 ttggcccggg ccttggagga tggatgttac acaatctgaa ttctttgaag tttacaaagg
     3421 ttgggatgaa gaagttttag atttgattcg gtgtgttgat aaacccacca aatgggcttt
     3481 acacgcttta gatcatttgg atgtttatgc gaaaggacga gtttttctta tgggggatgc
     3541 tgcccatgct atgttgcctc atttaggagc aggggctcat gttggaatgg aagatgcgta
     3601 tatcttggct tcattgatca ctcatagtag cacaccaatt tggccctcta cacaacacgt
     3661 tagtgaaatc gctaacatct acaacaccat gagaattcca agggccgttt caatgtccaa
     3721 ttctaccgat gaagcaggtt atttgtgcaa cttggaaaac cctggtttgg aagaattcaa
     3781 agttggagat catatcccaa aagaattgtt gatccaaaca gctagaacca tggagaaaaa
     3841 atgggcgtgg accactacat atgctgatga agatcggatt aaagccattt ccttattgga
     3901 aggtccccga gcagttttat aaaatcacac tcctaaaacc caaatcgtcc aaagaaaccg
     3961 tatagtatgc aactatagcc caaaacagac ccgatcctta atatggatct ccaacagata
     4021 ttgataaaat gccgattatc cttaacattt ttatctatgt ccatgagctg tcacgagcca
     4081 gccactggtc cctctagcgc ctgtatatca aagtatttta tggcagcaaa ttttaagttt
     4141 tgtaacggtt caatgcagat ctttacagag tttttcggct gatatgggct aaatccaaga
     4201 ttttataggg gttttaggaa ttcttaatta ttgttaccga ctcccccatg tgactcggaa
     4261 agatgatata caatttacag acaacagcgg ctgacggcag aatcaaaaac tgttgtagac
     4321 agaagcatac agttgtcgtt ataatatgcc tttaaatgcc caaggcaaaa ctaagggttt
     4381 gttgttgccc ccaacaaaag ttaagaacac tctcaacttt tgtgaaggat gatgaagggg
     4441 cttaaaaaat cccgtttcag aatctgcaac aatatttttg ttatttgtga tacggagagg
     4501 agagtctaaa tgactccaat cttgaaccat tacttcgcgc ggatcaattg gtccggggct
     4561 gccgcagtta atattgatac cttgcgagct ttgcatttaa aacataattg tactatcccc
     4621 tttgaaaact tggatgtttt gttgcctaga gaaatccaat tggataacca atctcccgaa
     4681 gaaaaattgg ttattgccag aagggggggt tactgctttg aacaaaacgg tgtttttgaa
     4741 cgggttttgc gagaattggg attcaacgtt cggtctttgt tggggcgagt tgttttgagt
     4801 aatccacccg cattacctcc aagaacacat aggttattgt tggttgaatt ggaagaagaa
     4861 aaatggatcg ccgatgttgg atttggaggg caaacattaa ccgcaccaat cagattggtt
     4921 tcagatttag ttcaaaccac tccccatggt gaatataggt tgttacaaga aggagatgat
     4981 tgggttttgc aattcaacca tcatcaacat tggcaatcaa tgtatagatt tgatttgtgt
     5041 gaacaacaac aatcagatta tgttatggga aacttctggt cagcgcattg gcctcaatcc
     5101 catttcagac atcatttgtt gatgtgcagg catttgccag atggtggaaa attgactttg
     5161 acaaacttcc atttcaccca ttatgaaaat ggtcatgctg ttgaacaaag aaatttgccc
     5221 gatgttgcct cattgtacgc agttatgcaa gaacaattcg ggttaggtgt tgatgatgcg
     5281 aaacatgggt ttacagttga tgaattggct ttagttatgg cggcttttga tacccatcct
     5341 gaagccggta aataacagat aatctagcag tttaggtttc agatggagtc agctagggtc
     5401 tttaacccca tcgtcaacta taagatggag agatggcata cggtgacact gttaaagcca
     5461 taactatatt gctcttgttg ataaaatcag aaactgaggc cacttgagag tattgtaaat
     5521 aggtagtaca gtagtgctac ctattttgtt ttaaaataac taattgagtt tatcgctggg
     5581 actatggttg ataatcagat gtcggaaaag cagaatattt tatcagcaga agctgatatt
     5641 tacttgcata aaagggatta tcaacaagca gccattttat atgaacaagc catcaatctc
     5701 aatcccgact ataaatataa ttactggaaa ctggggttaa ccctgctgct acaggagcgg
     5761 gaggaagaag cccagaccac ctggctggtg gcgatgagtg atggagaccc agaacagatt
     5821 gaccagtgga cgatagactt actcgaggtg ctatcatcag aagccgatgc acgaacctcc
     5881 gccgaagact attccgtggc ttgggggatt cgccaacaca ttttagaaat aaacccccat
     5941 ggtattgata atatattaaa gttaattgat ttgtcaatta ccctgacaac ctataccgga
     6001 aatgagatcg cagaatatgg actggttccc ttactgcaac aggagtccat caatgtcaat
     6061 ccagatttac tactgcacct gtggaaaaaa ctcctagttg tagatgcaat taatccccta
     6121 tctctggaat tgggagaggc ggtgattaac catcttggtg agcaaccgga gtttgtcact
     6181 agcttaatcc ctaaattata cgatattgcc tattcctttg gtgaaatagg tatagccaag
     6241 ggctatggag agttgttgtt gaacctggca ccgcatcatc aagagttgct gagggctctg
     6301 gctcaatttt gtattcaatt aaccgagtat gaccatggca ttgaatatgc cagactggcg
     6361 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6421 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6481 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6541 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6601 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6661 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6721 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6781 ctccgcctcc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7381 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7501 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7561 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7621 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7681 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7741 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7801 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7861 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7921 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7981 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8041 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8101 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8161 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8221 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8281 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8341 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8401 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8461 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8521 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8581 ccctttcgtc
//
