LOCUS       Exported                5275 bp ds-DNA     circular SYN 27-APR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 25, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..5275
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    14..143
                     /label=AAV2 ITR (alternate)
     repeat_region   25..52
                     /label=AAV2 ITR (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=19.9"
                     /note="/other=repeat_region"
     repeat_region   81..108
                     /label=AAV2 ITR (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=19.9"
                     /note="/other=repeat_region"
     promoter        177..624
                     /label=hSyn promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     primer_bind     602..621
                     /label=hSyn-F
     misc_feature    637..670
                     /label=FRT (minimal)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     intron          complement(671..720)
                     /label=beta-globin intron (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=10.5"
                     /note="/other=intron"
     misc_feature    721..754
                     /label=FRT5
     misc_feature    721..754
                     /label=FRT5
     misc_feature    767..800
                     /label=lox2272
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    851..884
                     /label=loxN
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    885..976
                     /label=T3H48 ribozyme
     ncRNA           924..973
                     /label=Hammerhead 1
                     /note="pLannotate"
                     /note="/database=Rfam"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=108.7"
                     /note="/other=ncRNA"
     protein_bind    complement(977..1010)
                     /label=lox2272
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     intron          complement(1011..1060)
                     /label=beta-globin intron (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=96.0"
                     /note="/match_length=10.5"
                     /note="/other=intron"
     protein_bind    complement(1061..1094)
                     /label=loxN
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     primer_bind     complement(1085..1109)
                     /label=EGFP-C
     CDS             complement(1109..1828)
                     /codon_start=1
                     /label=EGFP
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(1112..1828)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     1127..1145
                     /label=AH-Split-ATG-1209-F
     primer_bind     complement(1127..1145)
                     /label=AH-split-ATG-1209-R
     primer_bind     complement(1154..1175)
                     /label=EGFPCnew
     primer_bind     1186..1205
                     /label=eGFP_2
     primer_bind     complement(1689..1708)
                     /label=eGFP_1
     regulatory      1825..1834
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(1853..1886)
                     /label=FRT (minimal)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    complement(1937..1970)
                     /label=FRT5
     misc_feature    complement(1937..1970)
                     /label=FRT5
     primer_bind     complement(2011..2032)
                     /label=WPRE-5
     misc_feature    2021..2178
                     /label=WPRE (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=96.2"
                     /note="/match_length=39.1"
                     /note="/other=misc_feature"
     misc_feature    2022..2264
                     /label=wPRE3 (estimated)
     primer_bind     complement(2074..2094)
                     /label=WPRE_Rev
     misc_feature    2184..2268
                     /label=WPRE (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=14.4"
                     /note="/other=misc_feature"
     polyA_signal    complement(2267..2310)
                     /label=SV40 poly(A) signal (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=53.7"
                     /note="/other=polyA_signal"
     misc_feature    2267..2310
                     /label=SV40 late polyA
     misc_feature    2313..2447
                     /label=SL (estimated)
     polyA_signal    complement(2314..2381)
                     /label=SV40 poly(A) signal (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=82.9"
                     /note="/other=polyA_signal"
     misc_feature    2316..2359
                     /label=SV40 late polyA
     repeat_region   2467..2607
                     /label=AAV2 ITR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=repeat_region"
     repeat_region   2467..2596
                     /label=AAV2 ITR
     primer_bind     complement(2647..2663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2671..2687)
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     promoter        complement(2695..2725)
                     /label=lac promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     protein_bind    complement(2740..2761)
                     /label=CAP binding site
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     CDS             complement(2774..2866)
                     /codon_start=1
                     /label=lacI (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=8.6"
                     /note="/other=CDS"
                     /translation="APNTQTASPRALADSLMQLARQVSRLESGQ"
     rep_origin      complement(3049..3637)
                     /direction=LEFT
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             complement(3808..4668)
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.7"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4669..4773)
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             4868..4903
                     /codon_start=1
                     /label=rop (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=18.8"
                     /note="/other=CDS"
                     /translation="LARFGDDGENL"
     misc_feature    5005..5104
                     /label=bom (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=70.9"
                     /note="/other=misc_feature"
     CDS             complement(5110..5267)
                     /codon_start=1
                     /label=lacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=90.8"
                     /note="/other=CDS"
                     /translation="IHWPSFYNVVTGKTLALPNLIALQHIPLSPAGVIAKRPAPIALPN
                     SCAA*MA"
     primer_bind     5247..5263
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 attgggatcc aatcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
       61 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      121 caactccatc actaggggtt cctgcggccg cagatgctgc acgggcccgc gatcgcagtg
      181 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc
      241 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
      301 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
      361 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
      421 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
      481 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
      541 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
      601 gtcgtgtcgt gcctgagagc gcagggtacc gctagcgaag ttcctattct ctagaaagta
      661 taggaacttc gcagaatggt agctggattg tagctgctat tagcaatatg aaacctctta
      721 gaagttccta ttcttcaaaa ggtataggaa cttcgtcgac cttaagataa cttcgtatag
      781 gatactttat acgaagttat gtagctggag caatttgaaa agattgttgt attacctcta
      841 gctgcgctta ataacttcgt atagtatacc ttatacgaag ttattttctt tctttgcgcg
      901 tcctggattc gcggaaacgc gtacatccag ctgacgagtc ccaaatagga cgaaacgcgc
      961 ttttcttttt gtttttataa cttcgtataa agtatcctat acgaagttat ttgccttaac
     1021 ccagaaatta tcactgttat tctttagata ggtgcaaaga ataacttcgt ataaggtata
     1081 ctatacgaag ttatgaattc atgcatgctt acttgtacag ctcgtccatg ccgagagtga
     1141 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     1201 tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     1261 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     1321 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     1381 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     1441 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     1501 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     1561 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     1621 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     1681 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     1741 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     1801 tgaacagctc ctcgcccttg ctcaccatgg tggcactagt accggtctcg aggaagttcc
     1861 tatactttct agagaatagg aacttctaag cagacaatct tgtaatacat cagcagctag
     1921 tagattgctc cagctagaag ttcctatacc ttttgaagaa taggaacttc cgtacgaagc
     1981 tttctagagc ttagccgatt cgatattcga tgcccgagat aatcaacctc tggattacaa
     2041 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2101 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2161 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     2221 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta tttgtgaaat
     2281 ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt tgtgatgcta
     2341 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2401 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcggc cggtaaccgg
     2461 agctccagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2521 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2581 gcgagcgagc gcgcagctgc ctgcaggatt ggatcccaat ggcgcgccga gcttggcgta
     2641 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2701 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2761 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2821 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2881 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2941 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3001 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3061 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3121 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3181 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3241 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3301 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3361 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3421 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3481 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3541 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3601 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3661 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3721 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3781 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3841 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3901 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc
     3961 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4021 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4081 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4141 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4201 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4261 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4321 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4381 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4441 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4501 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4561 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4621 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4681 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4741 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4801 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4861 cttttgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4921 gactgtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4981 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5041 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5101 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5161 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5221 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattga cgcgt
//
