LOCUS       Exported                7276 bp ds-DNA     circular SYN 03-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7276)
  AUTHORS   Mingxi
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 5, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7276
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    2238..2555
                     /label=TetO
     protein_bind    2251..2521
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2554..2592
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    2556..2675
                     /label=miniCMV
     primer_bind     2600..2619
                     /label=PHAGE2-tetO-qPCR-F
     primer_bind     2655..2719
                     /label=GA-pHAGE2TetO-hSox2-F
     primer_bind     2657..2714
                     /label=hSOX2-MluI-AgeI-kozak-F
     primer_bind     complement(2665..2692)
                     /label=pHAGE2-Vector-AgeI-R
     regulatory      2679..2685
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     2685..2714
                     /label=hSOX2-AgeI-F
     CDS             2691..3650
                     /codon_start=1
                     /label=mSOX2-Anc_SoxCEF
                     /translation="MYNMMETELKPPGPQQASGGGGGGGNATAAATGGNQKNSPNHIKR
                     PMNAFMVWSQIERRKIAEQHPDMHNAEISKRLGKRWKLLSDDEKRPFVEEAERLRAQHM
                     QEHPDYKYRPRRKKKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGAGVNQRMDSYA
                     HMNGWSNGSYSMMQEQLGYPQHPGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGS
                     PTYSMSYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLP
                     GAEVPEPAAPSRLHMAQHYQSGPVPGTAINGTLPLSHM"
     misc_feature    2691..2705
                     /label=OXXOO
     primer_bind     complement(2703..2722)
                     /label=viral hSOX2-R
                     /note="DWH, IJSC, 2020"
     CDS             2811..3047
                     /codon_start=1
                     /label=Anc_SoxCEF HMG
                     /translation="NHIKRPMNAFMVWSQIERRKIAEQHPDMHNAEISKRLGKRWKLLS
                     DDEKRPFVEEAERLRAQHMQEHPDYKYRPRRKKK"
     misc_feature    3078..3092
                     /label=OXXOO
     misc_feature    3258..3272
                     /label=OXXOO
     misc_feature    3414..3428
                     /label=OXXOO
     misc_feature    3504..3530
                     /label=SOX17-bcat alignment
     misc_feature    3504..3518
                     /label=OXXOO
     misc_feature    3606..3620
                     /label=OXXOO
     primer_bind     complement(3623..3686)
                     /label=GA-pHAGE2TetO-hSox2-R
     primer_bind     3651..3683
                     /label=pHAGE2-Vector-SalI-F
     misc_feature    3670..4258
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4141..4152)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4256..4269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4388..4421
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     LTR             4454..4634
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        4767..4871
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4872..5732
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5090..5109)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5365..5374
                     /label=GA-pHAGE2TetO-hSox2-F
     rep_origin      5903..6491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6392..6411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6645..6662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6902..6923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6938..6968
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6976..6992
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7000..7016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca
     2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2461 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc caccaccggt atgtataaca
     2701 tgatggagac ggagctgaag ccgccgggcc cgcagcaagc ttcggggggc ggcggcggag
     2761 gaggcaacgc cacggcggcg gcaaccggcg gcaaccagaa gaacagcccg aatcacataa
     2821 aaaggccaat gaacgccttc atggtgtggt cccagatcga acggcgaaaa attgcagagc
     2881 aacatcctga catgcacaac gctgaaatct ctaaacggtt ggggaagcga tggaagttgc
     2941 tctctgacga cgagaaacga cccttcgtcg aggaggcaga gaggctcaga gctcaacaca
     3001 tgcaagagca tcctgactat aaatacagac caagacggaa gaagaagacg ctcatgaaga
     3061 aggataagta cacgcttccc ggaggcttgc tggcccccgg cgggaacagc atggcgagcg
     3121 gggttggggt gggcgccggc ctgggtgcgg gcgtgaacca gcgcatggac agctacgcgc
     3181 acatgaacgg ctggagcaac ggcagctaca gcatgatgca ggagcagctg ggctacccgc
     3241 agcacccggg cctcaacgct cacggcgcgg cacagatgca accgatgcac cgctacgacg
     3301 tcagcgccct gcagtacaac tccatgacca gctcgcagac ctacatgaac ggctcgccca
     3361 cctacagcat gtcctactcg cagcagggca cccccggtat ggcgctgggc tccatgggct
     3421 ctgtggtcaa gtccgaggcc agctccagcc cccccgtggt tacctcttcc tcccactcca
     3481 gggcgccctg ccaggccggg gacctccggg acatgatcag catgtacctc cccggcgccg
     3541 aggtgccgga gcccgctgcg cccagtagac tgcacatggc ccagcactac cagagcggcc
     3601 cggtgcccgg cacggccatt aacggcacac tgcccctgtc gcacatgtaa gtcgacatcg
     3661 atagatccta atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3721 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3781 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3841 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3901 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3961 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4021 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     4081 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4141 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4201 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4261 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     4321 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatcacc tgcaggacag
     4381 gcgcgccata acttcgtata gcatacatta tacgaagtta tggtacctgc gcgccctgct
     4441 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4501 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4561 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4621 gaaaatctct agcacccggg cgattaagga aagggctaga tcattcttga agacgaaagg
     4681 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4741 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4801 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4861 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4921 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4981 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5041 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5101 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5161 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5221 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5281 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5341 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5401 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5461 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5521 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5581 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5641 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5701 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5761 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5821 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5881 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5941 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6001 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6061 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6121 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6181 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6241 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6301 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6361 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6421 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6481 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6541 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6601 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6661 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6721 aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6781 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6841 aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6901 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6961 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     7021 ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7081 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
     7141 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
     7201 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
     7261 ctacaaactt agtagt
//
