LOCUS       sequence-51675-e       11002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11002)
  AUTHORS   Yoon HS, Golden JW
  TITLE     Heterocyst pattern formation controlled by a diffusible peptide.
  JOURNAL   Science. 1998 Oct 30;282(5390):935-8.
  PUBMED    9794762
REFERENCE   2  (bases 1 to 11002)
  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: "Science."; 
            date: "1998-10-30"; volume: "282(5390)"; pages: "935-8"
COMMENT     Sequence Label: pAM1954
FEATURES             Location/Qualifiers
     source          1..11002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      104..692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     593..612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     846..863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    878..1018
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(944..963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1104..1126
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1346..1365
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1571..2365)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1682..1701)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2292..2311
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             10259..10972
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(10289..10317)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 agctcgaatt gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
       61 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      241 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      481 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      721 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      781 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      841 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
      901 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
      961 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     1021 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     1081 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     1141 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     1201 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     1261 gcttctgata aagcgggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     1321 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     1381 ctgtggcgcc ggtgatgccg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     1441 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     1501 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     1561 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     1621 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     1681 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     1741 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     1801 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     1861 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     1921 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     1981 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     2041 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     2101 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     2161 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     2221 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     2281 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     2341 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc
     2401 ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
     2461 gcttcccaac cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata
     2521 aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg
     2581 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
     2641 gtttctgcgg actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt
     2701 gcttgcggca gcgtgaagct ttctctgagc tgtaacagcc tgaccgcaac aaacgagagg
     2761 atcgagacca tccgctccag attatccggc tcctccatgc gttgcctctc ggctcctgct
     2821 ccggttttcc atgccttatg gaactcctcg atccgccagc gatgggtata aatgtcgatg
     2881 acgcgcaagg cttgggctag cgactcgacc ggttcgccgg tcagcaacaa ccatttcaac
     2941 ggggtctcac ccttgggcgg gttaatctcc tcggccagca ccgcgttgag cgtgatattc
     3001 ccctgtttta gcgtgatgcg cccactgcgc atagaaattg catcaacgca tatagcgcta
     3061 gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatctaga cttatataga
     3121 cactaatata gacaatagtt tatactgcta tctatacaag tatagacatt atctaatcat
     3181 ggcagacaaa actctagcca cttttcgtat tgactccgaa gaatgggagt cttttaaaaa
     3241 ccttgctagt tctgaaagtt ccaacgcctc agcactgtta acagaatttg ttcgttggta
     3301 tttggcaggt aacaggttta atactcccac ttctcacact cccacccatc tagacacatc
     3361 cctcgaacag cgtatagaca atattgaaca acgtctagat aaagtcacaa ctaataatct
     3421 agacaatata gatgaattta tagacaagcg tatagaagat aatctagcaa cacgtctaga
     3481 caaacttcaa tcgcaactgg aggaactgcg gggaaaatcg aaagcccggt agttcaggca
     3541 gaaggacaag ctaccgggca agacagaaag aatatagaca atagtataga caatctagac
     3601 aaattggagg caacccgcga tcgcaccctc aataagctaa aaatgggtag gcagtcagcc
     3661 gccgggaaag ccatcgacgc gtttatcaaa gagttgcttt cttcaggaga caacataagc
     3721 tgaagttatc aaaattctgt ccttacgtcg aaagcctgat tttaccgtgc aacgattgat
     3781 aagcttggct aaactagcac tggctttcaa cagaaagcat acgaagaatc aatagatata
     3841 gccaccaatt ccacaaaatg cagataacgt gtagagtatt ggaatgctta atctgtaagg
     3901 gttatgaagg ttaacggcaa cggacgagcc aaaatactca cctccgacga actcaggcga
     3961 ctgtttagcg acggattcac cacaccgcgc gatcgcgttt tgtttggcat ctgtctattc
     4021 accggttgcc gcgttagtga agctctagca ctccaaacaa cggacattaa aggcgaaaca
     4081 ctaaccttta ggaagtctac caccaaaggg aaactcaaaa cccgcgtggt tgacatccag
     4141 ccaggactag ccgcactcat ggctgactat caccccaaac cgggaaccct gttccctggc
     4201 atgaggggag tcagcgatag gctcacgcga tacgcggcgg ataaaatctt gcgcgatgca
     4261 gccaaaagaa tcgggctaga aggcatcagt acccacagtt tccgccgtac tgccctcaac
     4321 caaatgtcta gcgccggtat cccgttgcga cacattcaag agatatccgg tcacaatgac
     4381 cttggcacac tgcaacgcta tcttgaagtt acacccgaac agcgacgcaa agctgtatcc
     4441 gtgattggct tctaatgtac gccaacgctg tttagacccc tatgggtgct aaaaaaagac
     4501 gcagcctaaa cacacgctct acacttgagg atacttttaa agtatccatc ggttctagaa
     4561 ctctgcacac gttccggact ttggaaacgt tatacctttc cctgtgttgc agaatgctgc
     4621 aatatttctt cgacaagtta acttgtgact ggtttaatat tttctcaaat tgccccaaaa
     4681 caacacgcct aaatccttag acgtttctgt ggaaacctat taggttttta tcgccgttgt
     4741 tttagtggta aacccaaagg gtttgtatat tcttgtatga agttcgactc tgagggttaa
     4801 gaagaatggc tcgccgaatt ttttacaagt ggaaaccgat taaaggttaa gggtcaatcg
     4861 ggacgatgaa tattttctaa ttgtgacctt ctccatctaa taagctttct ttggggttaa
     4921 ggtcgaagaa agtactacgc atgatctgca tacgatctct attgccaaaa agccgcgacc
     4981 ctataggctc tcggtcatgc tgcactagtt cgtgtcgatc actatactgg ttgccgcagc
     5041 atttcacgct aaaaaaaaat tcttaaaaat gtccttcata tctcgccaga gtggcaacct
     5101 attacaaaac ggttgcctac ccgaccggct cgattttcgc tgaagtggca ctgtgacagt
     5161 ttgaaatggt acttccgccg tgctgctgac atcgttgtta gggtgaattg ttcgcggtag
     5221 atgttgcacc gattcatgaa caccttgtca cccactttga ataatcgacc gtcaaattca
     5281 gtcgcgtcaa tttggtaagt gttgggctgt ctctttttgg ctccaggggc aatgccatca
     5341 gaaaacacaa ccgcgtcacc cataacttga taaccgatat cagttttggt tccagtgaaa
     5401 gcccaaaatt cagacgcgtc attattccga gcgtgccgga gttgattgta ctcaattttg
     5461 gcttggcaaa gttgacggcg attcatgccc agctgctttt gatgtcgtcg cactgtgcgc
     5521 ttgtgaatac ccaactcaca gctgacagct ttttgagatg taccatagtg gatgaaactt
     5581 tttgagacga atatccgcga cgaactaatg tgaagtacac aaggtacttc cccctctggc
     5641 gatttaagag aggattgcct tgtgtccttc actagctcgt tcgggtgtgg cgctccaaaa
     5701 agttttctgt actctggttt aagttgtctg ttggccgcat agcggctctt ttgttgaaag
     5761 ctttgtgtga ctatgccagt ggtcagtgag cgtaaatcgc ttaacacttg gactaaaggc
     5821 actactgcaa catcacccca tctttttaaa tttaggttgt aacaaacttg aaacataccg
     5881 cccaagtaga cggttatcat tcctgcttta attttgtagc ggcggaatgc tcctattttt
     5941 tttccatcct gtaaccaacg gtaaacagac ttatcactac aatctaagaa cgtctgtact
     6001 acaggcaatg gcaatgttaa atgaccagac ccatccttat caagcgctcg acacaaatac
     6061 cacaaccgcg cacaaggttc tcgaccaatg cgagtgtgta ccctgaccgt gtaagtgcca
     6121 agaattattt cagtttgtag ttcccttgta agcagggtta gtgatacatt tgtatttaag
     6181 ctttctgggc tgatcatttg gaaatgtctc agtccagtac ctattgaatg ttatttgctt
     6241 aacctgaagc taaataaaac ttgttaacta cacccattaa ttgataaatt caaagcacgt
     6301 tttttctgtt tggtgtttgg tgtggtaaca attctgtgta tgtgtgtttt atttagcttc
     6361 ggttaagtag cataacaacc tccaagcact gaactttttt taataggtaa tttaaacttt
     6421 gcctatcggc aaaattttca atcaattgta cgccaaagtg ttgcatgatc aacgtttgac
     6481 ttatttttgt atttactaaa tactgaattt cgccgtgacg ctttttacag atggaaattc
     6541 acggcaaaat gttttttgct aactttgcta tgtaaaacaa gaaacttggc actcggttat
     6601 tactaaataa actggtaaaa aataaccatt agaaccaaaa agaacgaaaa ccagtacacc
     6661 cttgccagtt ttcaagcttt tgctatgacg actctaataa tcgggtttaa caccattccg
     6721 ctttgagaaa attatccttg tacagcaagt aacagtcaat gctaaacgca ccgctacaaa
     6781 tccttaagtt tttccagtag cgatttacct tcttggtaac tgcccgcctt gatagcccag
     6841 aatgtcttta atcactgtaa ctttctgaaa aacgcctgcg ttccagacag gcttttgcca
     6901 cttttgctag ggtttcatct cttggatctg gtagggatga aacgggctgt aatgcttgtt
     6961 ctgaggtcgg ttgacgccgt tgtggacgtg ggtgaaaact ggttacagac tgaaccgggg
     7021 cataaccatt atgtaactgc tttacatctt ggtaactgac acggcatatc atccaccatg
     7081 cagcgatatt tccccgagct ttaaccactc cacaagggca aggtctttta aggacttggc
     7141 gtggctaact gcaaacttac ccaggcgtaa catcctaaac acttacggac accgccttca
     7201 ccctcgatac ctaaggtctt gacattatca tcttgagtca gcccaataac aaaacgcttg
     7261 ggcttgcggc cgcgcctggc gtgtttgatg agccattcgg ttgctatctc gacttcatct
     7321 ctcagcagtg gcagttcttc agcaattaaa acgctttctt ttcctgctag tgccttatcc
     7381 ccagactcac cccgtagctc aatccggcgc tgcaattcct ccaggtcagc agccatgccc
     7441 gactgtatag cctcaaagtc accacggcgg ccaatgacat ttaaccccgt ccactcgtcc
     7501 ggtgcagcgt cagcgtcata gactgtcacc tcacccccga cttgataagc aagccattgg
     7561 gctatggtgc ttttgccagt tcccgtatcc ccaactatta aacagtgctt accagacaga
     7621 gcttgcatca agtcggtgat gattccctct ggttcgaccg caagggtgac ggcggtagtg
     7681 tcaatgatag ccgcgccgta agtgccagca tagggcaatt ggtcgtaaac tttgaccaag
     7741 ttgtatacag actgtctaca ccacttcacc actgttaacg ctgtttgcaa agcgtaagac
     7801 gtggcatcaa ataaaaatat gctggcacta aaagttaatc gccccaatcc ccacagtaaa
     7861 aacctgccta gctgttgacg actaggcaag tgcatttcaa tccagtcatt tgccataaat
     7921 caccccgtct ttaaagcctt gcagttgagc gcgacaggta tttaactgtg cttgtaactc
     7981 tgtttgctgg ttttgatacc acagactgac ggcggcggcc gccagtccta aaaatagaaa
     8041 ctggcgatcg ctcattattg acttactccc tgttgattag cgtggtagtg agtcatagcc
     8101 gcattgaccg cttcttgggc ttggggtgtt ctgccaagat tgggttttgt agggtcatcg
     8161 ttggctacga ctaaggacgc ttgttcggct atcgcttgcg ggacaccaac tttagttaac
     8221 tctgtcaagg atacttggta aagtcgctcg ttcattagcc gattctccgg tacataaaac
     8281 tgttgctggc agtcccttca ttggcgacga gttcttcagc cggagtatca gcgataatgt
     8341 cagcccagcc ggtgacatta ttattaataa tgttttgttc ggcaattgca cccaagccag
     8401 gacgcgccgt ttcaaactca gagatgactt gctgctcttt ctcggtgagt ggtctatctg
     8461 tcatgataat tatgtccttc attatgtagg cgattccagt gggtgtttac gaggcagtcc
     8521 acaggaatca gtgcgattca cctttaaggt gaatcgtcat caaaaaatca ctcggtagca
     8581 acgacccgaa ccgaccagga ttgatttccc ggttctcagt tcgcaggctt ttgagcgcgt
     8641 caccttgacc attgggtaac tgccatcagc cgataagcta aacgggctgt atagcggtaa
     8701 agcatcccac acagtcgggc tggcatcaac tttgcaggaa tagctcacgt cactcatctc
     8761 actcgcgcct gggttggatg gcagcgaagg cagattacga cgcagttttt tactggcact
     8821 tttacccgca ttaaaaacgg gtacagtgcc attgttgacg gtctgtactt cggtcatata
     8881 ctcggtgtac acttaataca ctctatacta ttactgccga ttagtacatt tgtcaatcac
     8941 tctttgcaca aggtgtatga tatggactca ggagtacacc aaacgtcatg ccaaccaata
     9001 aagggagaat agcagtcact ctagaagctg aaatttacca atggattgct aaccgagcgt
     9061 ctgaggaagg aagaccgttg gctaatcttg ccgctttctt actcacacga gttgttaaag
     9121 aacaaatgga acaagaagcc aaggacaacc aagacaagca gggggcagca tgagcgaaga
     9181 cagactagcc agaatagaag ctgcgttaga cagccaagtt gcagtgaatg ccgacctccg
     9241 cacatcggtt acagaactcc gcgcaaccgc agaagcattg ttgcaaacag ttcaaatcca
     9301 tcagcagaac tttgaaattc ttaccgctag gcaattacaa accgaagcac ggcttgatga
     9361 gtaccaacgt accactagcg cggcactcga cagaattggc gcggtcttag actacctcgt
     9421 taggcagcaa aacggttgag gtgagggatg agcgatgact atctagacgg atatcccgca
     9481 agaggccctt tcgtcttcaa gaattcccgt ttgactggcg atgctgctac tgaaactaac
     9541 aactacatcg actacgcaat taacgccctc agctaatttt gcttagtcta ggcccggatg
     9601 ggtaagtggt tttcagctta agtgttgggt tctacttact tctccgggtc ttgctctatc
     9661 taaaaacatt ggtttaacaa ggagtattag gcaaatgcca gttactgtcg ctgcctctcg
     9721 cttgggaacc gctgcgtttg accaatcacc cgtcgaactg cgcgctaact attctcgacc
     9781 tgcaggtcga cggtatcgat aagcttgata acgcttttac tttttcctct tgccatagtt
     9841 aaaaaattct tttttatctg ttcttctcac ccactaaatt gcttttatcc cttcacgacc
     9901 ttcgcggttt acggaaagta tgagatgatt cccctatatt tcggggtatt taatgaattg
     9961 ggttagtttt cacacattcc cgcttctgca ctcttgccga cctcctttat tgacaggaga
    10021 cgagctaaga gccaatttac ttgacggata tttagtagag taatcagcgc aagatgaaaa
    10081 aattgcacac cacgtaattt gtaacttgtc tcacaatata tcaagaaatg gcataagtta
    10141 ttctttgaaa gttcttaact tacatattta aatttgttat tacataaagt tttaaacgtt
    10201 ttgttatcga attcctgcag cccgggggat cctctagatt taagaaggag atatacatat
    10261 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggcga
    10321 tgttaatggg caaaaattct ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
    10381 acttaccctt aaatttattt gcactactgg gaagctacct gttccatggc caacacttgt
    10441 cactactttc gcgtatggtc ttcaatgctt tgcgagatac ccagatcata tgaaacagca
    10501 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tattttacaa
    10561 agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa
    10621 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaaat
    10681 ggaatacaac tataactcac ataatgtata catcatggca gacaaaccaa agaatggaat
    10741 caaagttaac ttcaaaatta gacacaacat taaagatgga agcgttcaat tagcagacca
    10801 ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct
    10861 gtccacacaa tctgcccttt ccaaagatcc caacgaaaag agagatcaca tgatccttct
    10921 tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca aataagaatt
    10981 cagacctgca ggcatgcaag ct
//
