LOCUS       Exported                5921 bp ds-DNA     circular SYN 27-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5921)
  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..5921
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      928..1516
                     /label=ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             1699..1791
                     /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"
     protein_bind    1804..1825
                     /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"
     promoter        1840..1870
                     /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    1878..1894
                     /label=lac operator
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1958..2098
                     /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   1969..2098
                     /label=AAV2 ITR
     misc_feature    complement(2118..2252)
                     /label=SL (estimated)
     polyA_signal    2184..2251
                     /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    complement(2206..2249)
                     /label=SV40 late polyA
     polyA_signal    2255..2298
                     /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    complement(2255..2298)
                     /label=SV40 late polyA
     misc_feature    complement(2297..2381)
                     /label=WPRE (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=14.4"
                     /note="/other=misc_feature"
     misc_feature    complement(2301..2543)
                     /label=wPRE3 (estimated)
     misc_feature    complement(2387..2544)
                     /label=WPRE (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=96.2"
                     /note="/match_length=39.1"
                     /note="/other=misc_feature"
     protein_bind    2595..2642
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    2609..2642
                     /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    2705..2738
                     /label=FRT5
     misc_feature    2705..2738
                     /label=FRT5
     misc_feature    2762..2773
                     /label=present in sequenced plasmid
     misc_feature    2775..2780
                     /label=KOZAK
     CDS             2784..2810
                     /codon_start=1
                     /label=3xHA (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=33.3"
                     /note="/other=CDS"
                     /translation="YPYDVPDYA"
     CDS             2811..4250
                     /codon_start=1
                     /label=hM4D
                     /note="hM4D"
                     /translation="MANFTPVNGSSGNQSVRLVTSSSHNRYETVEMVFIATVTGSLSLV
                     TVVGNILVMLSIKVNRQLQTVNNYFLFSLACADLIIGAFSMNLYTVYIIKGYWPLGAVV
                     CDLWLALDCVVSNASVMNLLIISFDRYFCVTKPLTYPARRTTKMAGLMIAAAWVLSFVL
                     WAPAILFWQFVVGKRTVPDNQCFIQFLSNPAVTFGTAIAGFYLPVVIMTVLYIHISLAS
                     RSRVHKHRPEGPKEKKAKTLAFLKSPLMKQSVKKPPPGEAAREELRNGKLEEAPPPALP
                     PPPRPVADKDTSNESSSGSATQNTKERPATELSTTEATTPAMPAPPLQPRALNPASRWS
                     KIQIVTKQTGNECVTAIEIVPATPAGMRPAANVARKFASIARNQVRKKRQMAARERKVT
                     RTIFAILLAFILTWTPYNVMVLVNTFCQSCIPDTVWSIGYWLCYVNSTINPACYALCNA
                     TFKKTFRHLLLCQYRNIGTAR"
     CDS             2811..4247
                     /codon_start=1
                     /label=CHRM4
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=99.6"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MANFTPVNGSSGNQSVRLVTSSSHNRYETVEMVFIATVTGSLSLV
                     TVVGNILVMLSIKVNRQLQTVNNYFLFSLACADLIIGAFSMNLYTVYIIKGYWPLGAVV
                     CDLWLALDCVVSNASVMNLLIISFDRYFCVTKPLTYPARRTTKMAGLMIAAAWVLSFVL
                     WAPAILFWQFVVGKRTVPDNQCFIQFLSNPAVTFGTAIAGFYLPVVIMTVLYIHISLAS
                     RSRVHKHRPEGPKEKKAKTLAFLKSPLMKQSVKKPPPGEAAREELRNGKLEEAPPPALP
                     PPPRPVADKDTSNESSSGSATQNTKERPATELSTTEATTPAMPAPPLQPRALNPASRWS
                     KIQIVTKQTGNECVTAIEIVPATPAGMRPAANVARKFASIARNQVRKKRQMAARERKVT
                     RTIFAILLAFILTWTPYNVMVLVNTFCQSCIPDTVWSIGYWLCYVNSTINPACYALCNA
                     TFKKTFRHLLLCQYRNIGTAR"
     misc_feature    3650..3655
                     /label=KOZAK
     misc_feature    3656..3661
                     /label=KOZAK
     misc_feature    3711..3716
                     /label=KOZAK
     misc_feature    3762..3767
                     /label=KOZAK
     misc_feature    4182..4187
                     /label=KOZAK
     protein_bind    complement(4259..4292)
                     /label=loxP
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    4293..4384
                     /label=T3H48 ribozyme
     ncRNA           4332..4381
                     /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(4385..4418)
                     /label=loxP
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     protein_bind    complement(4431..4478)
                     /label=FRT
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=protein_bind"
     misc_feature    complement(4541..4574)
                     /label=FRT5
     misc_feature    complement(4541..4574)
                     /label=FRT5
     promoter        complement(4587..5034)
                     /label=hSyn promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     misc_feature    4748..4753
                     /label=KOZAK
     misc_feature    5068..5202
                     /label=AAV2 ITR (alternate)
     repeat_region   5103..5130
                     /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   5159..5186
                     /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"
     CDS             5219..5376
                     /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     complement(5223..5239)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(5382..5481)
                     /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(5583..5618)
                     /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    5689..5694
                     /label=KOZAK
     promoter        5713..5817
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     CDS             join(5818..5921,1..757)
                     /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"
ORIGIN
        1 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt ggttctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagctcggcg cgccattggg atccaatcct gcaggcagct gcgcgctcgc
     1981 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     2041 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgg
     2101 agctccggtt accggccgct ttaaaaaacc tcccacatct ccccctgaac ctgaaacata
     2161 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2221 gcaatagcat cacaaatttc acaaataaag ctagatggtt acaaataaag caatagcatc
     2281 acaaatttca caaataaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     2341 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     2401 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     2461 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     2521 aattttgtaa tccagaggtt gattatctcg ggcatcgaat atcgaatcgg ctaagctcta
     2581 gaaagcttcg tacggaagtt actattccga agttcctatt ctctagaaag tataggaact
     2641 tcgtgtcgat ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt cttcctattt
     2701 tgatgaagtt cctattcttc aaaaggtata ggaacttcct cgagaccggt atcgtggcta
     2761 gctctagagt cgacgccacc atgtacccat acgatgttcc agattacgct atggccaact
     2821 tcacacctgt caatggcagc tcgggcaatc agtccgtgcg cctggtcacg tcatcatccc
     2881 acaatcgcta tgagacggtg gaaatggtct tcattgccac agtgacaggc tccctgagcc
     2941 tggtgactgt cgtgggcaac atcctggtga tgctgtccat caaggtcaac aggcagctgc
     3001 agacagtcaa caactacttc ctcttcagcc tggcgtgtgc tgatctcatc ataggcgcct
     3061 tctccatgaa cctctacacc gtgtacatca tcaagggcta ctggcccctg ggcgccgtgg
     3121 tctgcgacct gtggctggcc ctggactgcg tggtgagcaa cgcctccgtc atgaaccttc
     3181 tcatcatcag ctttgaccgc tacttctgcg tcaccaagcc tctcacctac cctgcccggc
     3241 gcaccaccaa gatggcaggc ctcatgattg ctgctgcctg ggtactgtcc ttcgtgctct
     3301 gggcgcctgc catcttgttc tggcagtttg tggtgggtaa gcggacggtg cccgacaacc
     3361 agtgcttcat ccagttcctg tccaacccag cagtgacctt tggcacagcc attgctggct
     3421 tctacctgcc tgtggtcatc atgacggtgc tgtacatcca catctccctg gccagtcgca
     3481 gccgagtcca caagcaccgg cccgagggcc cgaaggagaa gaaagccaag acgctggcct
     3541 tcctcaagag cccactaatg aagcagagcg tcaagaagcc cccgcccggg gaggccgccc
     3601 gggaggagct gcgcaatggc aagctggagg aggccccccc gccagcgctg ccaccgccac
     3661 cgcgccccgt ggctgataag gacacttcca atgagtccag ctcaggcagt gccacccaga
     3721 acaccaagga acgcccagcc acagagctgt ccaccacaga ggccaccacg cccgccatgc
     3781 ccgcccctcc cctgcagccg cgggccctca acccagcctc cagatggtcc aagatccaga
     3841 ttgtgacgaa gcagacaggc aatgagtgtg tgacagccat tgagattgtg cctgccacgc
     3901 cggctggcat gcgccctgcg gccaacgtgg cccgcaagtt cgccagcatc gctcgcaacc
     3961 aggtgcgcaa gaagcggcag atggcggccc gggagcgcaa agtgacacga acgatctttg
     4021 ccattctgct tgccttcatc ctcacctgga cgccctacaa cgtcatggtc ctggtgaaca
     4081 ccttctgcca gagctgcatc cctgacacgg tgtggtccat tggctactgg ctctgctacg
     4141 tcaacagcac catcaaccct gcctgctatg ctctgtgcaa cgccaccttt aaaaagacct
     4201 tccggcacct gctgctgtgc cagtatcgga acatcggcac tgccaggtag ggcgcgccat
     4261 aacttcgtat aatgtatgct atacgaagtt attttctttc tttgcgcgtc ctggattcgc
     4321 ggaaacgcgt acatccagct gacgagtccc aaataggacg aaacgcgctt ttctttttgt
     4381 ttttataact tcgtataatg tatgctatac gaagttatct taaggtcgac gaagttccta
     4441 tactttctag agaataggaa cttcggaata ggaacttcgt gtcgatggtg atgcttggca
     4501 attcgggtcg atggtgaagc attggtcttc ctattttgat gaagttccta taccttttga
     4561 agaataggaa cttcgctagc ggtaccctgc gctctcaggc acgacacgac tcctccgctg
     4621 cccaccgcag actgaggcag cgctgagtcg ccggcgccgc agcgcagatg gtcgcgcccg
     4681 tgccccccta tctcgcgcct cgcgtggtgc ggtccggctg ggccggcggc ggcgcggacg
     4741 cgaccaaggt ggccgggaag gggagtttgc gggggaccgg cgagtgacgt cagcgcgcct
     4801 tcagtgctga ggcggcggtg gcgcgcgccg ccaggcgggg gcgaaggcac tgtccgcggt
     4861 gctgaagctg gcagtgcgca cgcgcctcgc cgcatcctgt ttcccctccc cctctctgat
     4921 aggggatgcg caatttgggg aatgggggtt gggtgcttgt ccagtgggtc ggggtcggtc
     4981 gtcaggtagg cacccccacc ccgcctcatc ctggtcctaa aacccacttg cactgcgatc
     5041 gcgggcccgt gcagcatctg cggccgcagg aacccctagt gatggagttg gccactccct
     5101 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg
     5161 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggatt ggatcccaat acgcgtcaat
     5221 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5281 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5341 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     5401 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     5461 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5521 gcttgtctgc tcccggcatc cgcttacaga caagctgtga cagtctccgg gagctgcatg
     5581 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac aaaagggcct cgtgatacgc
     5641 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5701 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5761 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5821 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5881 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg a
//
