LOCUS       pKG2899_pGEEC550        3553 bp    DNA     circular SYN 05-MAR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3553)
  AUTHORS   MIT
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..42
                     /label=oMJ001
     primer_bind     1..42
                     /label=oMJ005
     primer_bind     3..42
                     /label=oMJ009
     primer_bind     3..42
                     /label=oMJ013
     primer_bind     9..39
                     /label=oMJ020
     primer_bind     complement(21..46)
                     /label=oSEQ033 Pentr_rev
     misc_feature    23..46
                     /label=pENTR2B.Seq.REV
     primer_bind     complement(35..56)
                     /label=oTA069 mIR-backbone_junction_rev
     primer_bind     complement(35..56)
                     /label=oSEQ044 pENTR_rev_seq
     primer_bind     complement(108..128)
                     /label=oSEQ005 pENTR_rev
                     /note="25nm	STD"
     CDS             122..931
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(147..168)
                     /label=oSEQ023 attl2_rev
     primer_bind     complement(155..178)
                     /label=Kan_rev_SEQ
     primer_bind     complement(545..574)
                     /label=oTA099 GG_entry_upstream_fwd
     primer_bind     545..574
                     /label=oTA102 GG_entry_downstream_rev
     misc_feature    1005..1624
                     /label=pBR322_Origin
     rep_origin      1021..1609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1416..1431
                     /label=oTA073 Xho_addition_rev
     primer_bind     complement(1486..1501)
                     /label=oTA072 Xho_addition_for
     primer_bind     1510..1529
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     1761..1774
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      1774..1860
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1923..1942
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     terminator      1952..1979
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1981..2000
                     /label=pENTR_fwd_seq
     primer_bind     1981..2000
                     /label=oSEQ043 pENTR_fwd_seq
     primer_bind     complement(2002..2046)
                     /label=oMJ002
     primer_bind     complement(2002..2042)
                     /label=oMJ010
     primer_bind     complement(2002..2038)
                     /label=oMJ006
     primer_bind     complement(2002..2038)
                     /label=oMJ014
     primer_bind     complement(2010..2038)
                     /label=oMJ019
     primer_bind     2043..2069
                     /label=pPV1-CMV_fwd
     primer_bind     2043..2064
                     /label=oEP143
     enhancer        2047..2350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2218..2237)
                     /label=oSEQ065 CMV_rev
     promoter        2351..2559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2505..2525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2508..2528
                     /label=oSEQ030 CMV_fwd
     primer_bind     complement(2523..2542)
                     /label=oSEQ042 SFCp_rev
     primer_bind     complement(2536..2574)
                     /label=pPV1-CMV_rev
     primer_bind     complement(2540..2559)
                     /label=oEP138
     primer_bind     complement(2540..2559)
                     /label=oEP144
     primer_bind     2555..2572
                     /label=pPV1-EF1a-bb_fwd
     primer_bind     complement(2586..2610)
                     /label=pPV1-EF1a_rev
     primer_bind     2605..2637
                     /label=oKL145
     misc_feature    2610..2618
                     /label=Kozak sequence
     CDS             2616..3317
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(2616..2633)
                     /label=oKL095
     primer_bind     2717..2734
                     /label=?oSEQ052 Univ_FP_Fwd
     primer_bind     complement(2717..2734)
                     /label=oSEQ053 Univ_FP_Rev
     primer_bind     2880..2902
                     /label=oSEQ069 TagBFP_fwd
     primer_bind     complement(3009..3030)
                     /label=oSEQ070 TagBFP_rev
     primer_bind     complement(3179..3188)
                     /label=oSEQ011 pEntr_for
                     /note="25nm	STD"
     primer_bind     3229..3252
                     /label=oKL143
     primer_bind     complement(3229..3252)
                     /label=oKL144
     primer_bind     complement(3295..3321)
                     /label=oKL142
     primer_bind     3317..3341
                     /label=gebGH_fwd
     primer_bind     3323..3344
                     /label=GG_bGH_fwd
     primer_bind     3327..3344
                     /label=GG_WPRE-bGH_fwd
     polyA_signal    3329..3553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(join(3535..3553,1..4))
                     /label=gebGH_rev
     primer_bind     complement(3539..3553)
                     /label=GG_bGH_rev
ORIGIN
        1 cgctcgctga ctgcaggtgc ccgtgtctca aaatctctga tgttacattg cacaagataa
       61 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
      121 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
      181 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
      241 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
      301 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
      361 gaccatcaag cattttatcc gtactcctgg tgatgcatgg ttactcacca ctgcgatccc
      421 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
      481 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
      541 cagcgatcgc gtatttcgtc tagctcaggc gcaatcacga atgaataacg gtttggttga
      601 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
      661 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
      721 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
      781 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
      841 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
      901 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta
      961 ttcagattgg gccccgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1141 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1681 gttatcccct gattctgtgg ataaccgtat taccgctagc atggatctcg gggacgtcta
     1741 actactaagc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
     1801 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1861 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
     1921 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
     1981 gcgtttctac aaactcttcc tgttagttag ttacttaagc tcgggcccac ctgcagtcta
     2041 cttactcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2101 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2161 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2221 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2281 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2341 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2401 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2461 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2521 gcgtgtacgg tgggaggtct atataagcag agctgagctc gtttagtgaa ccgtcagatc
     2581 gcctggagac gccatccacg ctgtaatgag ccaccatgag cgagctgatt aaggagaaca
     2641 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     2701 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     2761 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     2821 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     2881 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     2941 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     3001 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc
     3061 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     3121 atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga
     3181 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaaa
     3241 cgtacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     3301 ggcacaagct taattaacaa cgttgcacct gtgccttcta gttgccagcc atctgttgtt
     3361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3541 gtgggctcta tgg
//
