LOCUS       Exported                3603 bp ds-DNA     circular SYN 06-MAR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3603)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 14, 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..3603
                     /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
     misc_feature    703..732
                     /locus_tag="SGSGGSGGSG linker"
                     /label=SGSGGSGGSG linker
     gene            733..1449
                     /locus_tag="mGFP"
                     /label=mGFP
     primer_bind     complement(796..815)
                     /locus_tag="oSC186 reverse"
                     /label=oSC186 reverse
     misc_feature    1453..1475
                     /locus_tag="3' homology arm"
                     /label=3' homology arm
     primer_bind     complement(join(1453..1458,1467..1468))
                     /locus_tag="Reverse primer"
                     /label=Reverse primer
     misc_feature    1469
                     /locus_tag="PCR insertion between TA(1)"
                     /label=PCR insertion between TA
                     /label=PCR insertion between TA(1)
     misc_recomb     complement(1485..1505)
                     /locus_tag="attB2"
                     /label=attB2
     misc_feature    complement(1492..1498)
                     /locus_tag="Gateway Recomb2"
                     /label=Gateway Recomb2
     primer_bind     complement(1598..1619)
                     /locus_tag="T7"
                     /label=T7
     primer_bind     complement(1618..1638)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     promoter        1716..1849
                     /locus_tag="SpnR Promoter"
                     /label=SpnR Promoter
     gene            1850..2860
                     /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 ccagtggttc cggaggatct
      721 ggaggatctg gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      781 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      841 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      901 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      961 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1021 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1081 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1141 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1201 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1261 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1321 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1381 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1441 ctgtacaagt agcatatggc ggccgcggaa gggcgaattc gacccagctt tcttgtacaa
     1501 agttggcatt ataaaaaata attgctcatc aatttgttgc aacgaacagg tcactatcag
     1561 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1621 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1681 acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac
     1741 ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     1801 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     1861 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     1921 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     1981 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     2041 cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac
     2101 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     2161 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     2221 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     2281 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     2341 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     2401 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     2461 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     2521 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     2581 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     2641 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     2701 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     2761 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     2821 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     2881 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     2941 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3001 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3061 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3121 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     3361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3541 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3601 gtt
//
