LOCUS       Exported                4533 bp DNA     circular SYN 01-DEC-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4533)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4533)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4533)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(131..164)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     oriT            377..486
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      601..989
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1022..2068)
                     /codon_start=1
                     /label=mariner transposase C9 mutant
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     promoter        complement(2092..2122)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2137..2158)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    2182..2210
                     /label=MmeI IR_R
                     /note="MmeI modified inverted repeats"
     terminator      complement(2256..2283)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(2375..2461)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2520..3314)
                     /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"
     misc_feature    3667..3695
                     /label=MmeI IR_L
                     /note="MmeI modified inverted repeats"
     protein_bind    complement(3732..3779)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(3796..4455)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(join(4456..4533,1..25))
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
       61 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
      121 taggcgcgcc gaagttccta tactttctag agaataggaa cttcggaata ggaactaagg
      181 aggatattca catggaccat ggctaattcc catacgcgtc aattcgaggg ggatcaattc
      241 cgttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat
      301 cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta
      361 tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct
      421 tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct
      481 ggccggctac cgccggcgta acagatgagg gcaagcggat ggctgatgaa accaagccaa
      541 ccaggaaggg cagcccacct atcacggaat tgatccggcc acgatgcgtc cggcgtagag
      601 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
      661 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
      721 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct
      781 catgtttgaa caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag
      841 ttttataaga aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg
      901 gacaacaagc cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg
      961 agtgacacag gaacacttaa cggctgacat gggaattccc ctccaccgcg gtggcggccg
     1021 cttattcaac atagttccct tcaagagcga tacaacgatt ataacgacct tccaattttt
     1081 tgataccatt ttggtagtac tccttcggtt ttgcctcaaa ataggcctca gtttcggcga
     1141 tcacctcttc attgcagcca aattttttcc ctgcgagcat ccttttgagg tctgagaaca
     1201 agaaaaagtc gctgggggcc agatctggag aatacggcgg gtggggaagc aattcgaagc
     1261 ccaattcatg aatttttgcc atcgttctca atgacttgtg gcacggtgcg ttgtcttggt
     1321 ggaacaacac ttttttcttc ttcatgtggg gccgttttgc cgcgatttcg accttcaaac
     1381 gctccaataa cgccatataa tagtcactgt tgatggtttt tcccttctca agataatcga
     1441 taaaaattat tccatgcgca tcccaaaaaa cagaggccat tactttgcca gcggactttt
     1501 gagtctttcc acgcttcgga gacggttcac cggtcgctgt ccactcagcc gactgtcgat
     1561 tggactcagg agtgtagtga tggagccatg tttcatccat tgtcacatat cgacggaaaa
     1621 actcgggtgt attacgagtt aacagctgca aacaccgctt agaatcatca acacgtcgtt
     1681 gtttttggtc aaatgtgagc tcgcgcggca cccatttcgc acagagcttc cgcatatcca
     1741 aatattgatg aatgatatga ccaacacgtt cctttgatat ctttaaggcc tctgctatct
     1801 cgatcaactt cattttacgg tcattcaaaa tcattttgtg gatttttttg atgttttcgt
     1861 cggtaaccac ctctttcggg cgtccactgc gttcaccgtc ctccgtgctc atttcaccac
     1921 gcttgaattt tgcataccaa tcaattattg ttgatttccc tggggcagag tccggaaact
     1981 cattatcaag ccaagttttt gcttccactg tattttttcc cttcagaaaa cagtatttta
     2041 tcaaaacacg aaattccttt ttttccatat gtatccgctc acaattccac acaacatacg
     2101 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2161 tgcgttgcgc tcactggatc ctaacaggtt ggatgataag tccccggtct tcgtatgccg
     2221 tcttctgctt ggcgcgccct gcaggtagaa acgcaaaaag gccatccgtc aggatggcct
     2281 tctgcttaat ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     2341 gttgcttcgc aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     2401 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
     2461 gatgcctggc agttccctac tctcgcattc tagagtcatt tcgaacccca gagtcccgct
     2521 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     2581 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     2641 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     2701 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     2761 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     2821 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     2881 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     2941 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     3001 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     3061 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     3121 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     3181 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     3241 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     3301 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct tgatcccctg
     3361 cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc
     3421 ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag
     3481 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     3541 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     3601 ctggctttct acgtgttccg cttcctcaat tgctcgaggg cgcgccaagc agaagacggc
     3661 atacgaagac cggggactta tcatccaacc tgttaggtac cgaggacgcg tgtgtaggct
     3721 ggagctgctt cgaagttcct atactttcta gagaatagga acttcggaat aggaacttca
     3781 tttaaatggc gcgccttacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     3841 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     3901 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     3961 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     4021 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     4081 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     4141 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     4201 ccatatcacc agctcaccgt ctttcattgc catacgtaat tccggatgag cattcatcag
     4261 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     4321 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     4381 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     4441 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacaact caaaaaatac
     4501 gcccggtagt gatcttattt cattatggtg aaa
//
