LOCUS       Exported                6276 bp ds-DNA     circular SYN 06-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 13, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     For adding mGFP to C-terminal of proteins using KpnI site and Gibson
            cloning.
FEATURES             Location/Qualifiers
     source          1..6276
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     536..553
                     /locus_tag="M13-fwd"
                     /label=M13-fwd
     misc_recomb     570..665
                     /locus_tag="AttL1"
                     /label=AttL1
     misc_recomb     645..665
                     /locus_tag="attB1"
                     /label=attB1
     misc_feature    652..658
                     /locus_tag="Gateway recomb1"
                     /label=Gateway recomb1
     misc_feature    670..696
                     /locus_tag="5' homology arm"
                     /label=5' homology arm
     misc_feature    682
                     /locus_tag="PCR insertion between TA"
                     /label=PCR insertion between TA
     primer_bind     join(683..684,691..696)
                     /locus_tag="Forward primer"
                     /label=Forward primer
     CDS             703..3369
                     /locus_tag="tiam-1"
                     /label=tiam-1
     misc_feature    1906..1925
                     /locus_tag="Overlap for gibson"
                     /label=Overlap for gibson
     misc_feature    3376..3405
                     /locus_tag="SGSGGSGGSG linker"
                     /label=SGSGGSGGSG linker
     gene            3406..4122
                     /locus_tag="mGFP"
                     /label=mGFP
     primer_bind     complement(3469..3488)
                     /locus_tag="oSC186 reverse"
                     /label=oSC186 reverse
     misc_feature    4126..4148
                     /locus_tag="3' homology arm"
                     /label=3' homology arm
     primer_bind     complement(join(4126..4131,4140..4141))
                     /locus_tag="Reverse primer"
                     /label=Reverse primer
     misc_feature    4142
                     /locus_tag="PCR insertion between TA(1)"
                     /label=PCR insertion between TA
                     /label=PCR insertion between TA(1)
     misc_recomb     complement(4158..4178)
                     /locus_tag="attB2"
                     /label=attB2
     misc_feature    complement(4165..4171)
                     /locus_tag="Gateway Recomb2"
                     /label=Gateway Recomb2
     primer_bind     complement(4271..4292)
                     /locus_tag="T7"
                     /label=T7
     primer_bind     complement(4291..4311)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     promoter        4389..4522
                     /locus_tag="SpnR Promoter"
                     /label=SpnR Promoter
     gene            4523..5533
                     /locus_tag="SpnR"
                     /label=SpnR
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gaattcgccc ttggactagt catatgggta ccatgggctc acgcctctca
      721 tgttcttgct cacaagccga atgctggacg aatgatgaat ctacattgag aattgacgcg
      781 gagttgtttg agttatcaca ggacggacaa aagtgggata agcttgacgg aaggctggcc
      841 aacgtacgtg tattcaacgc attcgacgac tcttccccac gtcttcttgc cacatcatca
      901 tccggtcaag ttcttctgga cactctaatt ccaatgggag aaaaggttca taaagtcagc
      961 gacttttttg tgtatttgaa gacggaagga agaacaattg gattcaatac gttaagctcg
     1021 agggacactt ccttactcat ttctcaggcg acaaatacaa ctgccttcaa tatgtttcaa
     1081 cattcattct cagtttccgt atccagaatt gctacatttg aaggtgattc tattcaacct
     1141 tgtgatgagc agccagaaac gtcaaatgtc tcactaaatc cgttaaatat ggttttcaac
     1201 tacaaaggat cacctgttga gattgatttg ctggaatgtt tcgcgtgctc atcagaagct
     1261 gaaaactgtg tcgacatagc acatgccaac aacatatttc gtgtaattgt gtcagcacat
     1321 gctcatcttt ttcttattca agctctgaat gtatcatcac ttcttcttgc tcgaaccacc
     1381 tcgaatctag ttgccatcaa ttatttggct cggcaaaatg ataaaattca acaaaaattg
     1441 attgctctgt tggaaacatc agaatcaagt gagaatgcgt cgaaggaaga tattcgacaa
     1501 atgcttcttc tgaagaatgt agtagtgaca cagagattga actcacttca acagagagca
     1561 ctaccgggaa ttgagcttat ccaacacgat atctcacctc aaattgatca gaagctactc
     1621 gaaaagtgtc gacttccatc actgttttta tatcatttta atatgaattc gtttcgaata
     1681 tggcgaaaac caatggaaac acgtgacatg aacacaatgg gatcatcaat ggcatccacc
     1741 agtcaatacc atcagattcc gattcaaatc cagaatcaga atccgttacc gtctcaacag
     1801 attcagccac aacaacaaca acaataccat ccaccagatc atgtgacaac tcataatggt
     1861 acaactacgt ttcatactac gaatcccaca aaaccaagac tttgtgaagg atcgttggct
     1921 cctggatgct cattggccga ggcatctgaa cagagaaaac gagagatggc ggagcagaaa
     1981 aaagagttgg caggcccatc ttccgcagag gattacttcg tcagaaagac aaatggacgt
     2041 cttggactca ctatatacgc gcataacgat gatggggtca tcagggctga ggttcgtgga
     2101 gtgacgtcat ttgctccgag atgtgctcaa gttggagatt cggttgtagc ggtagacagt
     2161 gaactcatat catcagttag aaacgcttcg gatgttgaga aacttttgag aatcggaaaa
     2221 gtaatacatt tacgaagaaa gactcctcta actccaagag caccgtctca actcgacgca
     2281 actaatgtca ataagaaaac gggctgtgca aagttggcga tggctctgca agaattgttg
     2341 gtcactgaga agaaatatgt cagcgatctt cgagagatga atgaagtttt tctgtgtatg
     2401 agacaagtcc gagatattat gaatgcagca atacggttgt ataaaatgca aacatcgttt
     2461 gtggacagta ttgaggaagc cattggagac atgagtagac aggatatttc tgttgctcag
     2521 attagggatt ctgtaatgcg agtgtgtgca gtattcatca acaaatgttc tgatttcaaa
     2581 atctatgcag aatatgcagc cggttaccat ctcctccaac atgagatcaa aaacaaaaaa
     2641 gagcttctat caaaattgga agctgtaaac agtacaagag agcaacattg ttcttgggaa
     2701 tctcggatga ttaaaccagt ccagagaatt gtacaatatc cacttttact gaaaaacatt
     2761 gccgatgcac ttccgaagga tgcacgagag agggttcaag tggaggcagc attgcaaaaa
     2821 atgcagacgt ctgcagaata tgtgaatgaa atgcaaaggc ttcatgaaga ttatgcacaa
     2881 tatatggaga ctgttaggaa agctaatgag ccaatgttgg ctgaaaagag tcttcgcctc
     2941 gacactcgag agcttctagt ttttgcacac ataaaatgga gagatgctcc agcggagcac
     3001 tacgttgtgt tcgtgtttca ctctttaata ctgctccttc catcgtatgc taggaaagag
     3061 attaagatga aatggactcg tgtcctccca ataaatgaag ttgatatcaa cgaaatgcca
     3121 aatgatgcat taaatctaaa actttatcat gctgcttttg aaggaccaaa tggacaatta
     3181 agtcattcca atccaaatac agtctacaat attgaatgtt gtcaatctca actgaaacag
     3241 catcttatca aaaatatcaa aaaagctcga actttattct ctcgagaatc acatcgtcca
     3301 ttaagtggat ctagtcaatc ggatggtggt tatgtgtcag aagtctctaa ggaacatcga
     3361 aattcaaaag gtaccagtgg ttccggagga tctggaggat ctggtatggt gagcaagggc
     3421 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3481 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3541 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     3601 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     3661 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3721 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3781 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3841 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     3901 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3961 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4021 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4081 accgccgccg ggatcactct cggcatggac gagctgtaca agtagcatat ggcggccgcg
     4141 gaagggcgaa ttcgacccag ctttcttgta caaagttggc attataaaaa ataattgctc
     4201 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     4261 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     4321 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
     4381 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctgcccaagg ttgccgggtg
     4441 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     4501 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     4561 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     4621 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     4681 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     4741 catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     4801 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     4861 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     4921 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     4981 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     5041 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     5101 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     5161 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     5221 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     5281 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     5341 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     5401 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     5461 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     5521 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
     5581 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5641 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5701 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5761 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5821 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5881 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5941 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6001 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6061 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6121 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6181 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6241 gttcctggcc ttttgctggc cttttgctca catgtt
//
