LOCUS       sequence-377057-        4636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains homology arms for the NSI (NIES39_Q01230) 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: pJMH021
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 ccagtggtgc tgtggcgatc gatggcttag
      421 aaatgctgat tcaacaagga gccgccgccc tggaaatttg gctacaacaa cccatcccga
      481 tctcagtcat gcgtcaatct ttaatcattg ataattgatc attcataatt gagctataat
      541 cgtgggggag ggtttggctg ttaattgtca attccccagg aatgttttgt tgaacctgcc
      601 cctggggaat ggtgcagtag taccgacaga ctaagaccct agcaagtatt tttaggtaca
      661 atttacatat taagcattaa tactctgggg atgaatttat gataacccta tatattgtct
      721 gtctaacggt gggtggagta tgtgtcatat tagccgctgt ggagggatta gacggtgtag
      781 agtttgaggg cgaatttgat agcgatgtag aattagaaca tggcaggggt gagaccgaaa
      841 accagcccag ctatagatcc aaaagaaaaa atctgcggtt aaggctgcct tttgggagtg
      901 tgcgcttctg gacttttggg gtgtgctttt ttgggttgac cggaacaatt atgtcagttt
      961 tgaaccccga cttaaccccg gcttttgtgg ctacagcagc gatcgctatt ggggtgtcct
     1021 gcggaaccgc tgtcgtgtgg gtctttgata ttctgcgatc gcaacaagct aacagtttag
     1081 ttcaatccca tgatttaatt ggtctatggg ccaccgtgga aattcccttt aatcgagaca
     1141 gtcgtggaaa agtacgcctc agcgccggag ggtcggtgct gtatttgagc gcaattactg
     1201 aagaaaatca ggaattttcc cagggcgatc gcgtttttgt ggtaggaatg gagaaaaata
     1261 aagttttggt ggtctcagaa gcatccttcg gagtctcaga agcataatcg acaagtccca
     1321 agtcagggta agcggtagta gagatccgac atttcacctt ttaaatgttt gctcaaaccc
     1381 ttttggggga aaaactctag atcaatttga tggcaaccaa acacaggtac aggaattatg
     1441 gaaattatca tcgccttatt agcagccctg ggtttaggaa ccggagcgac tgtatttatt
     1501 atccgcaatc tctactatat ttgtcagcct agcgaagtgt tgatttttgc cggaggtcgt
     1561 cgccctacac aagatgatcg cactgtggga tatcgcctgg tgaaaggtgg tagcagcatt
     1621 cggacacccc tgctagaaca agcctttcac atggatttaa ccaatatgat tattgagtta
     1681 aaggtggcta atgcttactc gaaaggaggc attcctttac aggttgatgg ggtggcgaat
     1741 attaaaattg ctggggaaga acccacaatt cataatgcga tcgagcgttt gttggggaaa
     1801 actcgccagg aaattgaacg cattgctaaa gataccctgg aaggaaacct gcgcggggta
     1861 ttagccagcc ttactcctga acaggttaat ggggataaat tggcttttgc taaaagtctc
     1921 ctcgaagaag ctgaggatga cttagaacaa ctggggctga ttttagatac tctccaaatt
     1981 caaaatattt ctgatgacgt gggatatctt gactccatcg gacgacaaca acgggcggaa
     2041 ttgtttagag attctcgtgt cgctgaagca caagctaagg ctgatgctgc tatccgtact
     2101 gcggaaaatc ataaaatgac tcagttgaaa aagttggaaa ctgaggtaga ggtatctaag
     2161 gcggaagctg aacgccgggt ggctgatgct atgactaaac gggcggcggt ggtagcagag
     2221 tcggagtcgg aaacagcggc tgaggtggct aggactcagg ctgaggtctc tgttcaaaaa
     2281 gaaaggatta agcaggtaga acagcaacta caagctgatg tggtcgcacc agcagaggcg
     2341 gaatgtaaaa aggcgatcgc tagggcgcgg ggggatgctg cccaaattat tgaagatggg
     2401 atggcgtaat 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
//
