LOCUS       sequence-377060-        4636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains homology arms for the pilA (NIES39_C03030) locus of 
            Arthrospira platensis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4636)
  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 4636)
  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: pJMH020
FEATURES             Location/Qualifiers
     source          1..4636
                     /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(2415..2431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2415..2431)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2428..2450)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2439..2455
                     /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(2463..2469,2470..2487,2488..2493))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2508..2529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2646..2663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2817..3405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2897..2916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3576..4367,4368..4436))
                     /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     4199..4218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4437..4541)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4609..4627
                     /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 cgaattctag acagataatc tagcagttta ggtttcagat ggagtcagct
     1441 agggtcttta accccatcgt caactataag atggagagat ggcatacggt gacactgtta
     1501 aagccataac tatattgctc ttgttgataa aatcagaaac tgaggccact tgagagtatt
     1561 gtaaataggt agtacagtag tgctacctat tttgttttaa aataactaat tgagtttatc
     1621 gctgggacta tggttgataa tcagatgtcg gaaaagcaga atattttatc agcagaagct
     1681 gatatttact tgcataaaag ggattatcaa caagcagcca ttttatatga acaagccatc
     1741 aatctcaatc ccgactataa atataattac tggaaactgg ggttaaccct gctgctacag
     1801 gagcgggagg aagaagccca gaccacctgg ctggtggcga tgagtgatgg agacccagaa
     1861 cagattgacc agtggacgat agacttactc gaggtgctat catcagaagc cgatgcacga
     1921 acctccgccg aagactattc cgtggcttgg gggattcgcc aacacatttt agaaataaac
     1981 ccccatggta ttgataatat attaaagtta attgatttgt caattaccct gacaacctat
     2041 accggaaatg agatcgcaga atatggactg gttcccttac tgcaacagga gtccatcaat
     2101 gtcaatccag atttactact gcacctgtgg aaaaaactcc tagttgtaga tgcaattaat
     2161 cccctatctc tggaattggg agaggcggtg attaaccatc ttggtgagca accggagttt
     2221 gtcactagct taatccctaa attatacgat attgcctatt cctttggtga aataggtata
     2281 gccaagggct atggagagtt gttgttgaac ctggcaccgc atcatcaaga gttgctgagg
     2341 gctctggctc aattttgtat tcaattaacc gagtatgacc atggcattga atatgccaga
     2401 ctggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2461 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2521 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2581 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3001 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3241 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4561 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4621 acgaggccct ttcgtc
//
