LOCUS       sequence-51676-e       10604 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 10604)
  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 10604)
  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: pAM1956
FEATURES             Location/Qualifiers
     source          1..10604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      107..695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     596..615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     849..866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    881..1021
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(947..966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1107..1129
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1349..1368
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1574..2368)
                     /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(1685..1704)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2295..2314
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             9858..10571
                     /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(9888..9916)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 ggggatcctc taggatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
       61 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      181 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      241 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      481 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      721 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      781 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      841 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
      901 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
      961 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     1021 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     1081 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     1141 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     1201 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     1261 ctggcttctg ataaagcggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     1321 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     1381 cacctgtggc gccggtgatg ccggtgggcg aagaactcca gcatgagatc cccgcgctgg
     1441 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     1501 gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac
     1561 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     1621 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     1681 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     1741 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     1801 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     1861 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     1921 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     1981 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     2041 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     2101 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     2161 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     2221 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     2281 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     2341 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     2401 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     2461 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc
     2521 ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct
     2581 ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc
     2641 accgtttctg cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg
     2701 agtgcttgcg gcagcgtgaa gctttctctg agctgtaaca gcctgaccgc aacaaacgag
     2761 aggatcgaga ccatccgctc cagattatcc ggctcctcca tgcgttgcct ctcggctcct
     2821 gctccggttt tccatgcctt atggaactcc tcgatccgcc agcgatgggt ataaatgtcg
     2881 atgacgcgca aggcttgggc tagcgactcg accggttcgc cggtcagcaa caaccatttc
     2941 aacggggtct cacccttggg cgggttaatc tcctcggcca gcaccgcgtt gagcgtgata
     3001 ttcccctgtt ttagcgtgat gcgcccactg cgcatagaaa ttgcatcaac gcatatagcg
     3061 ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatct agacttatat
     3121 agacactaat atagacaata gtttatactg ctatctatac aagtatagac attatctaat
     3181 catggcagac aaaactctag ccacttttcg tattgactcc gaagaatggg agtcttttaa
     3241 aaaccttgct agttctgaaa gttccaacgc ctcagcactg ttaacagaat ttgttcgttg
     3301 gtatttggca ggtaacaggt ttaatactcc cacttctcac actcccaccc atctagacac
     3361 atccctcgaa cagcgtatag acaatattga acaacgtcta gataaagtca caactaataa
     3421 tctagacaat atagatgaat ttatagacaa gcgtatagaa gataatctag caacacgtct
     3481 agacaaactt caatcgcaac tggaggaact gcggggaaaa tcgaaagccc ggtagttcag
     3541 gcagaaggac aagctaccgg gcaagacaga aagaatatag acaatagtat agacaatcta
     3601 gacaaattgg aggcaacccg cgatcgcacc ctcaataagc taaaaatggg taggcagtca
     3661 gccgccggga aagccatcga cgcgtttatc aaagagttgc tttcttcagg agacaacata
     3721 agctgaagtt atcaaaattc tgtccttacg tcgaaagcct gattttaccg tgcaacgatt
     3781 gataagcttg gctaaactag cactggcttt caacagaaag catacgaaga atcaatagat
     3841 atagccacca attccacaaa atgcagataa cgtgtagagt attggaatgc ttaatctgta
     3901 agggttatga aggttaacgg caacggacga gccaaaatac tcacctccga cgaactcagg
     3961 cgactgttta gcgacggatt caccacaccg cgcgatcgcg ttttgtttgg catctgtcta
     4021 ttcaccggtt gccgcgttag tgaagctcta gcactccaaa caacggacat taaaggcgaa
     4081 acactaacct ttaggaagtc taccaccaaa gggaaactca aaacccgcgt ggttgacatc
     4141 cagccaggac tagccgcact catggctgac tatcacccca aaccgggaac cctgttccct
     4201 ggcatgaggg gagtcagcga taggctcacg cgatacgcgg cggataaaat cttgcgcgat
     4261 gcagccaaaa gaatcgggct agaaggcatc agtacccaca gtttccgccg tactgccctc
     4321 aaccaaatgt ctagcgccgg tatcccgttg cgacacattc aagagatatc cggtcacaat
     4381 gaccttggca cactgcaacg ctatcttgaa gttacacccg aacagcgacg caaagctgta
     4441 tccgtgattg gcttctaatg tacgccaacg ctgtttagac ccctatgggt gctaaaaaaa
     4501 gacgcagcct aaacacacgc tctacacttg aggatacttt taaagtatcc atcggttcta
     4561 gaactctgca cacgttccgg actttggaaa cgttatacct ttccctgtgt tgcagaatgc
     4621 tgcaatattt cttcgacaag ttaacttgtg actggtttaa tattttctca aattgcccca
     4681 aaacaacacg cctaaatcct tagacgtttc tgtggaaacc tattaggttt ttatcgccgt
     4741 tgttttagtg gtaaacccaa agggtttgta tattcttgta tgaagttcga ctctgagggt
     4801 taagaagaat ggctcgccga attttttaca agtggaaacc gattaaaggt taagggtcaa
     4861 tcgggacgat gaatattttc taattgtgac cttctccatc taataagctt tctttggggt
     4921 taaggtcgaa gaaagtacta cgcatgatct gcatacgatc tctattgcca aaaagccgcg
     4981 accctatagg ctctcggtca tgctgcacta gttcgtgtcg atcactatac tggttgccgc
     5041 agcatttcac gctaaaaaaa aattcttaaa aatgtccttc atatctcgcc agagtggcaa
     5101 cctattacaa aacggttgcc tacccgaccg gctcgatttt cgctgaagtg gcactgtgac
     5161 agtttgaaat ggtacttccg ccgtgctgct gacatcgttg ttagggtgaa ttgttcgcgg
     5221 tagatgttgc accgattcat gaacaccttg tcacccactt tgaataatcg accgtcaaat
     5281 tcagtcgcgt caatttggta agtgttgggc tgtctctttt tggctccagg ggcaatgcca
     5341 tcagaaaaca caaccgcgtc acccataact tgataaccga tatcagtttt ggttccagtg
     5401 aaagcccaaa attcagacgc gtcattattc cgagcgtgcc ggagttgatt gtactcaatt
     5461 ttggcttggc aaagttgacg gcgattcatg cccagctgct tttgatgtcg tcgcactgtg
     5521 cgcttgtgaa tacccaactc acagctgaca gctttttgag atgtaccata gtggatgaaa
     5581 ctttttgaga cgaatatccg cgacgaacta atgtgaagta cacaaggtac ttccccctct
     5641 ggcgatttaa gagaggattg ccttgtgtcc ttcactagct cgttcgggtg tggcgctcca
     5701 aaaagttttc tgtactctgg tttaagttgt ctgttggccg catagcggct cttttgttga
     5761 aagctttgtg tgactatgcc agtggtcagt gagcgtaaat cgcttaacac ttggactaaa
     5821 ggcactactg caacatcacc ccatcttttt aaatttaggt tgtaacaaac ttgaaacata
     5881 ccgcccaagt agacggttat cattcctgct ttaattttgt agcggcggaa tgctcctatt
     5941 ttttttccat cctgtaacca acggtaaaca gacttatcac tacaatctaa gaacgtctgt
     6001 actacaggca atggcaatgt taaatgacca gacccatcct tatcaagcgc tcgacacaaa
     6061 taccacaacc gcgcacaagg ttctcgacca atgcgagtgt gtaccctgac cgtgtaagtg
     6121 ccaagaatta tttcagtttg tagttccctt gtaagcaggg ttagtgatac atttgtattt
     6181 aagctttctg ggctgatcat ttggaaatgt ctcagtccag tacctattga atgttatttg
     6241 cttaacctga agctaaataa aacttgttaa ctacacccat taattgataa attcaaagca
     6301 cgttttttct gtttggtgtt tggtgtggta acaattctgt gtatgtgtgt tttatttagc
     6361 ttcggttaag tagcataaca acctccaagc actgaacttt ttttaatagg taatttaaac
     6421 tttgcctatc ggcaaaattt tcaatcaatt gtacgccaaa gtgttgcatg atcaacgttt
     6481 gacttatttt tgtatttact aaatactgaa tttcgccgtg acgcttttta cagatggaaa
     6541 ttcacggcaa aatgtttttt gctaactttg ctatgtaaaa caagaaactt ggcactcggt
     6601 tattactaaa taaactggta aaaaataacc attagaacca aaaagaacga aaaccagtac
     6661 acccttgcca gttttcaagc ttttgctatg acgactctaa taatcgggtt taacaccatt
     6721 ccgctttgag aaaattatcc ttgtacagca agtaacagtc aatgctaaac gcaccgctac
     6781 aaatccttaa gtttttccag tagcgattta ccttcttggt aactgcccgc cttgatagcc
     6841 cagaatgtct ttaatcactg taactttctg aaaaacgcct gcgttccaga caggcttttg
     6901 ccacttttgc tagggtttca tctcttggat ctggtaggga tgaaacgggc tgtaatgctt
     6961 gttctgaggt cggttgacgc cgttgtggac gtgggtgaaa actggttaca gactgaaccg
     7021 gggcataacc attatgtaac tgctttacat cttggtaact gacacggcat atcatccacc
     7081 atgcagcgat atttccccga gctttaacca ctccacaagg gcaaggtctt ttaaggactt
     7141 ggcgtggcta actgcaaact tacccaggcg taacatccta aacacttacg gacaccgcct
     7201 tcaccctcga tacctaaggt cttgacatta tcatcttgag tcagcccaat aacaaaacgc
     7261 ttgggcttgc ggccgcgcct ggcgtgtttg atgagccatt cggttgctat ctcgacttca
     7321 tctctcagca gtggcagttc ttcagcaatt aaaacgcttt cttttcctgc tagtgcctta
     7381 tccccagact caccccgtag ctcaatccgg cgctgcaatt cctccaggtc agcagccatg
     7441 cccgactgta tagcctcaaa gtcaccacgg cggccaatga catttaaccc cgtccactcg
     7501 tccggtgcag cgtcagcgtc atagactgtc acctcacccc cgacttgata agcaagccat
     7561 tgggctatgg tgcttttgcc agttcccgta tccccaacta ttaaacagtg cttaccagac
     7621 agagcttgca tcaagtcggt gatgattccc tctggttcga ccgcaagggt gacggcggta
     7681 gtgtcaatga tagccgcgcc gtaagtgcca gcatagggca attggtcgta aactttgacc
     7741 aagttgtata cagactgtct acaccacttc accactgtta acgctgtttg caaagcgtaa
     7801 gacgtggcat caaataaaaa tatgctggca ctaaaagtta atcgccccaa tccccacagt
     7861 aaaaacctgc ctagctgttg acgactaggc aagtgcattt caatccagtc atttgccata
     7921 aatcaccccg tctttaaagc cttgcagttg agcgcgacag gtatttaact gtgcttgtaa
     7981 ctctgtttgc tggttttgat accacagact gacggcggcg gccgccagtc ctaaaaatag
     8041 aaactggcga tcgctcatta ttgacttact ccctgttgat tagcgtggta gtgagtcata
     8101 gccgcattga ccgcttcttg ggcttggggt gttctgccaa gattgggttt tgtagggtca
     8161 tcgttggcta cgactaagga cgcttgttcg gctatcgctt gcgggacacc aactttagtt
     8221 aactctgtca aggatacttg gtaaagtcgc tcgttcatta gccgattctc cggtacataa
     8281 aactgttgct ggcagtccct tcattggcga cgagttcttc agccggagta tcagcgataa
     8341 tgtcagccca gccggtgaca ttattattaa taatgttttg ttcggcaatt gcacccaagc
     8401 caggacgcgc cgtttcaaac tcagagatga cttgctgctc tttctcggtg agtggtctat
     8461 ctgtcatgat aattatgtcc ttcattatgt aggcgattcc agtgggtgtt tacgaggcag
     8521 tccacaggaa tcagtgcgat tcacctttaa ggtgaatcgt catcaaaaaa tcactcggta
     8581 gcaacgaccc gaaccgacca ggattgattt cccggttctc agttcgcagg cttttgagcg
     8641 cgtcaccttg accattgggt aactgccatc agccgataag ctaaacgggc tgtatagcgg
     8701 taaagcatcc cacacagtcg ggctggcatc aactttgcag gaatagctca cgtcactcat
     8761 ctcactcgcg cctgggttgg atggcagcga aggcagatta cgacgcagtt ttttactggc
     8821 acttttaccc gcattaaaaa cgggtacagt gccattgttg acggtctgta cttcggtcat
     8881 atactcggtg tacacttaat acactctata ctattactgc cgattagtac atttgtcaat
     8941 cactctttgc acaaggtgta tgatatggac tcaggagtac accaaacgtc atgccaacca
     9001 ataaagggag aatagcagtc actctagaag ctgaaattta ccaatggatt gctaaccgag
     9061 cgtctgagga aggaagaccg ttggctaatc ttgccgcttt cttactcaca cgagttgtta
     9121 aagaacaaat ggaacaagaa gccaaggaca accaagacaa gcagggggca gcatgagcga
     9181 agacagacta gccagaatag aagctgcgtt agacagccaa gttgcagtga atgccgacct
     9241 ccgcacatcg gttacagaac tccgcgcaac cgcagaagca ttgttgcaaa cagttcaaat
     9301 ccatcagcag aactttgaaa ttcttaccgc taggcaatta caaaccgaag cacggcttga
     9361 tgagtaccaa cgtaccacta gcgcggcact cgacagaatt ggcgcggtct tagactacct
     9421 cgttaggcag caaaacggtt gaggtgaggg atgagcgatg actatctaga cggatatccc
     9481 gcaagaggcc ctttcgtctt caagaattcc cgtttgactg gcgatgctgc tactgaaact
     9541 aacaactaca tcgactacgc aattaacgcc ctcagctaat tttgcttagt ctaggcccgg
     9601 atgggtaagt ggttttcagc ttaagtgttg ggttctactt acttctccgg gtcttgctct
     9661 atctaaaaac attggtttaa caaggagtat taggcaaatg ccagttactg tcgctgcctc
     9721 tcgcttggga accgctgcgt ttgaccaatc acccgtcgaa ctgcgcgcta actattctcg
     9781 acctgcaggt cgactctaga ggatcaattc gagctcggta cccggggatc ctctagattt
     9841 aagaaggaga tatacatatg agtaaaggag aagaactttt cactggagtt gtcccaattc
     9901 ttgttgaatt agatggcgat gttaatgggc aaaaattctc tgtcagtgga gagggtgaag
     9961 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactggg aagctacctg
    10021 ttccatggcc aacacttgtc actactttcg cgtatggtct tcaatgcttt gcgagatacc
    10081 cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg
    10141 aaagaactat attttacaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg
    10201 aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa
    10261 acattcttgg acacaaaatg gaatacaact ataactcaca taatgtatac atcatggcag
    10321 acaaaccaaa gaatggaatc aaagttaact tcaaaattag acacaacatt aaagatggaa
    10381 gcgttcaatt agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
    10441 taccagacaa ccattacctg tccacacaat ctgccctttc caaagatccc aacgaaaaga
    10501 gagatcacat gatccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
    10561 aactatacaa ataagaattc agacctgcag gcatgcaagc tggg
//
