LOCUS       Exported                6744 bp ds-DNA     circular SYN 05-DEC-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 22, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..6744
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(261..365)
                     /label=AmpR promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     rep_origin      complement(391..846)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(418..846)
                     /direction=LEFT
                     /label=f1 ori
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=rep_origin"
     CDS             complement(851..1006)
                     /codon_start=1
                     /label=lacZ-alpha (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=89.7"
                     /note="/other=CDS"
                     /translation="SLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQ
                     LRSLNGE"
     primer_bind     988..1004
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1014..1032
                     /label=T7 promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     intron          complement(1130..1195)
                     /label=small t intron
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=intron"
     CDS             complement(1197..1269)
                     /codon_start=1
                     /label=small t antigen (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=13.9"
                     /note="/other=CDS"
                     /translation="SL*RNLTSVV*HNWTNYLQRFKAL"
     CDS             complement(1272..1844)
                     /codon_start=1
                     /label=DTA
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     promoter        complement(1863..1990)
                     /label=HSV TK promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=87.7"
                     /note="/other=promoter"
     misc_feature    complement(2132..2157)
                     /label=MCS (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=24.1"
                     /note="/other=misc_feature"
     misc_feature    2163..3105
                     /label=SOX2-Homology arm left
     CDS             2164..3105
                     /codon_start=1
                     /label=SOX2
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=99.7"
                     /note="/match_length=99.1"
                     /note="/other=CDS"
                     /translation="MMETELKPPGPQQTSGGGGGNSTAAAAGGNQKNSPDRVKRPMNAF
                     MVWSRGQRRKMAQENPKMHNSEISKRLGAEWKLLSETEKRPFIDEAKRLRALHMKEHPD
                     YKYRPRRKTKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGAGVNQRMDSYAHMNGW
                     SNGSYSMMQDQLGYPQHPGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSM
                     SYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLPGAEVP
                     EPAAPSRLHMSQHYQSGPVPGTAINGTLPLSHM"
     CDS             3121..3177
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3184..3903
                     /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"
     misc_feature    3904..4902
                     /label=Sox2 homology arm right
     misc_feature    complement(4911..4930)
                     /label=MCS (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=18.5"
                     /note="/other=misc_feature"
     promoter        complement(4946..4962)
                     /label=T3 promoter (fragment)
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=89.5"
                     /note="/other=promoter"
     primer_bind     complement(4983..4999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5007..5023)
                     /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(5031..5061)
                     /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(5076..5097)
                     /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(5110..5202)
                     /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(5385..5973)
                     /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(join(6144..6744,1..260))
                     /codon_start=1
                     /label=AmpR
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=99.8"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
       61 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
      121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
      181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
      241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
      301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      361 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
      421 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
      481 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
      541 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
      601 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
      661 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
      721 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
      781 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
      841 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      901 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      961 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     1021 actcactata gggcgaattg ggtaccgggc cccccctcga ggtccccctc gactctagtg
     1081 gatctgcatt ccaccactgc tcccattcat cagttccata ggttggaatc taaaatacac
     1141 aaacaattag aatcagtagt ttaacacatt atacacttaa aaattttata tttaccttag
     1201 agctttaaat ctctgtaggt agtttgtcca attatgtcac accacagaag taaggttcct
     1261 tcacaaagag atcgcctgac acgatttcct gcacaggctt gagccatata ctcatacatc
     1321 gcatcttggc cacgttttcc acgggtttca aaattaatct caagttctac gcttaacgct
     1381 ttcgcctgtt cccagttatt aatatattca acgctagaac tcccctcagc gaagggaagg
     1441 ctgagcacta cacgcgaagc accatcaccg aaccttttga taaactcttc cgttccgact
     1501 tgctccatca acggttcagt gagacttaaa cctaactctt tcttaatagt ttcggcatta
     1561 tccactttta gtgcgagaac cttcgtcagt cctggatacg tcactttgac cacgcctcca
     1621 gcttttccag agagcgggtt ttcattatct acagagtatc ccgcagcgtc gtatttattg
     1681 tcggtactat aaaacccttt ccaatcatcg tcataatttc cttgtgtacc agattttggc
     1741 ttttgtatac ctttttgaat ggaatctaca taaccaggtt tagtcccgtg gtacgaagaa
     1801 aagttttcca tcacaaaaga tttagaagaa tcaacaacat catcaggatc catggcgagg
     1861 acctgcaggg tcgctcggtg ttcgaggcca cacgcgtcac cttaatatgc gaagtggacc
     1921 tgggaccgcg ccgccccgac tgcatctgcg tgttcgaatt cgccaatgac aagacgctgg
     1981 gcggggtttg ctcgacattg ggtggaaaca ttccaggcct gggtggagag gctttttgct
     2041 tcctcttgca aaaccacact gctcgacatt gggtggaaac attccaggcc tgggtggaga
     2101 ggctttttgc ttcctcttga aaaccacact gcgacggtat cgataagctt gatatcgtcg
     2161 accatgatgg agacggagct gaagccgccg ggcccgcagc aaacttcggg gggcggcggc
     2221 ggcaactcca ccgcggcggc ggccggcggc aaccagaaaa acagcccgga ccgcgtcaag
     2281 cggcccatga atgccttcat ggtgtggtcc cgcgggcagc ggcgcaagat ggcccaggag
     2341 aaccccaaga tgcacaactc ggagatcagc aagcgcctgg gcgccgagtg gaaacttttg
     2401 tcggagacgg agaagcggcc gttcatcgac gaggctaagc ggctgcgagc gctgcacatg
     2461 aaggagcacc cggattataa ataccggccc cggcggaaaa ccaagacgct catgaagaag
     2521 gataagtaca cgctgcccgg cgggctgctg gcccccggcg gcaatagcat ggcgagcggg
     2581 gtcggggtgg gcgccggcct gggcgcgggc gtgaaccagc gcatggacag ttacgcgcac
     2641 atgaacggct ggagcaacgg cagctacagc atgatgcagg accagctggg ctacccgcag
     2701 cacccgggcc tcaatgcgca cggcgcagcg cagatgcagc ccatgcaccg ctacgacgtg
     2761 agcgccctgc agtacaactc catgaccagc tcgcagacct acatgaacgg ctcgcccacc
     2821 tacagcatgt cctactcgca gcagggcacc cctggcatgg ctcttggctc catgggttcg
     2881 gtggtcaagt ccgaggccag ctccagcccc cctgtggtta cctcttcctc ccactccagg
     2941 gcgccctgcc aggccgggga cctccgggac atgatcagca tgtatctccc cggcgccgag
     3001 gtgccggaac ccgccgcccc cagcagactt cacatgtccc agcactacca gagcggcccg
     3061 gtgcccggca cagccattaa cggcacactg cccctctcac acatggtcga cggaagcgga
     3121 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctctc
     3181 gacatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3241 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3301 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3361 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3421 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3481 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3541 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3601 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3661 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3721 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3781 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3841 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3901 taagggccgg acagcgaact ggagggggga gaaattttca aagaaaaacg agggaaatgg
     3961 gaggggtgca aaagaggaga gtaagaaaca gcatggagaa aacccggtac gctcaaaaag
     4021 aaaaaggaaa aaaaaaatcc catcacccac agcaaatgac agctgcaaaa gagaacacca
     4081 atcccatcca cactcacgca aaaaccgcga tgccgacaag aaaactttta tgagagagat
     4141 cctggacttc tttttggggg actatttttg tacagagaaa acctggggag ggtggggagg
     4201 gcgggggaat ggaccttgta tagatctgga ggaaagaaag ctacgaaaaa ctttttaaaa
     4261 gttctagtgg tacggtagga gctttgcagg aagtttgcaa aagtctttac caataatatt
     4321 tagagctagt ctccaagcga cgaaaaaaat gttttaatat ttgcaagcaa cttttgtaca
     4381 gtatttatcg agataaacat ggcaatcaaa atgtccattg tttataagct gagaatttgc
     4441 caatattttt caaggagagg cttcttgctg aattttgatt ctgcagctga aatttaggac
     4501 agttgcaaac gtgaaaagaa gaaaattatt caaatttgga cattttaatt gtttaaaaat
     4561 tgtacaaaag gaaaaaatta gaataagtac tggcgaacca tctctgtggt cttgtttaaa
     4621 aagggcaaaa gttttagact gtactaaatt ttataactta ctgttaaaag caaaaatggc
     4681 catgcaggtt gacaccgttg gtaatttata atagcttttg ttcgatccca actttccatt
     4741 ttgttcagat aaaaaaaacc atgaaattac tgtgtttgaa atattttctt atggtttgta
     4801 atatttctgt aaatttattg tgatatttta aggttttccc ccctttattt tccgtagttg
     4861 tattttaaaa gattcggctc tgtattattt gaatcagtct gcgtttaaac ccgccaccgc
     4921 ggtggagctc cagcttttgt tcccttttag tgagggttaa ttgcgcgctt ggcgtaatca
     4981 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5041 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5101 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6721 tagtgtatgc ggcgaccgag ttgc
//
