LOCUS       Exported                7561 bp ds-DNA     circular SYN 02-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7561)
  AUTHORS   Mingxi
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 31, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7561
                     /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    2251..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"
     primer_bind     2554..2571
                     /label=pFlag-CMV-F
     misc_feature    2556..2675
                     /label=miniCMV
     primer_bind     2589..2614
                     /label=pLNCX-F
     regulatory      2679..2685
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2691..3935
                     /codon_start=1
                     /label=hSOX17
                     /translation="MSSPDAGYASDDQSQTQSALPAVMAGLGPCPWAESLSPIGDMKVK
                     GEAPANSGAPAGAAGRAKGESRIRRPMNAFMVWAKDERKRLAQQNPDLHNAELSKMLGK
                     SWKALTLAEKRPFVEEACRLRVQHMQDHPNYKYRPRRRKQVKRLKRVEGGFLHGLAEPQ
                     AAALGPEGGRVAMDGLGLQFPEQGFPAGPPLLPPHMGGHYRDCQSLGAPPLDGYPLPTP
                     DTSPLDGVDPDPAFFAAPMPGDCPAAGTYSYAQVSDYAGPPEPPAGPMHPRLGPEPAGP
                     SIPGLLAPPSALHVYYGAMGSPGAGGGRGFQMQPQHQHQHQHQHHPPGPGQPSPPPEAL
                     PCRDGTDPSQPAELLGEVDRTEFEQYLHFVCKPEMGLPYQGHDSGVNLPDSHGAISSVV
                     SDASSAVYYCNYPDV"
     CDS             2886..3122
                     /codon_start=1
                     /label=HMG
                     /translation="SRIRRPMNAFMVWAKDERKRLAQQNPDLHNAELSKMLGKSWKALT
                     LAEKRPFVEEACRLRVQHMQDHPNYKYRPRRRKQ"
     misc_feature    3027..3029
                     /label=Silent mutE48
     primer_bind     complement(3035..3066)
                     /label=hSOXE57R-R-3
     misc_feature    3048..3050
                     /label=Silent mutE55
     misc_feature    3054..3056
                     /label=E57C
     misc_feature    3955..4543
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4008..4028)
                     /label=WPRE-R
     primer_bind     4541..4554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4673..4706
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     LTR             4739..4919
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5052..5156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5157..6017
                     /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(5375..5394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6188..6776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6677..6696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6930..6947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7187..7208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7223..7253
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7261..7277
                     /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     7285..7301
                     /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 atgagcagcc
     2701 cggatgcggg atacgccagt gacgaccaga gccagaccca gagcgcgctg cccgcggtga
     2761 tggccgggct gggcccctgc ccctgggccg agtcgctgag ccccatcggg gacatgaagg
     2821 tgaagggcga ggcgccggcg aacagcggag caccggccgg ggccgcgggc cgagccaagg
     2881 gcgagtcccg tatccggcgg ccgatgaacg ctttcatggt gtgggctaag gacgagcgca
     2941 agcggctggc gcagcagaat ccagacctgc acaacgccga gttgagcaag atgctgggca
     3001 agtcgtggaa ggcgctgacg ctggcggaaa agcggccctt cgtggaggaa gcatgccggc
     3061 tgcgcgtgca gcacatgcag gaccacccca actacaagta ccggccgcgg cggcgcaagc
     3121 aggtgaagcg gctgaagcgg gtggagggcg gcttcctgca cggcctggct gagccgcagg
     3181 cggccgcgct gggccccgag ggcggccgcg tggccatgga cggcctgggc ctccagttcc
     3241 ccgagcaggg cttccccgcc ggcccgccgc tgctgcctcc gcacatgggc ggccactacc
     3301 gcgactgcca gagtctgggc gcgcctccgc tcgacggcta cccgttgccc acgcccgaca
     3361 cgtccccgct ggacggcgtg gaccccgacc cggctttctt cgccgccccg atgcccgggg
     3421 actgcccggc ggccggcacc tacagctacg cgcaggtctc ggactacgct ggccccccgg
     3481 agcctcccgc cggtcccatg cacccccgac tcggcccaga gcccgcgggt ccctcgattc
     3541 cgggcctcct ggcgccaccc agcgcccttc acgtgtacta cggcgcgatg ggctcgcccg
     3601 gggcgggcgg cgggcgcggc ttccagatgc agccgcaaca ccagcaccag caccagcacc
     3661 agcaccaccc cccgggcccc ggacagccgt cgccccctcc ggaggcactg ccctgccggg
     3721 acggcacgga ccccagtcag cccgccgagc tcctcgggga ggtggaccgc acggaatttg
     3781 aacagtatct gcacttcgtg tgcaagcctg agatgggcct cccctaccag gggcatgact
     3841 ccggtgtgaa tctccccgac agccacgggg ccatttcctc ggtggtgtcc gacgccagct
     3901 ccgcggtata ttactgcaac tatcctgacg tgtgagtcga catcgataga tcctaatcaa
     3961 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4021 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4081 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4141 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4201 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4261 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4321 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4381 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4441 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4501 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc
     4561 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     4621 agggctaatt cactcccaac gaagacaaga tcacctgcag gacaggcgcg ccataacttc
     4681 gtatagcata cattatacga agttatggta cctgcgcgcc ctgctttttg cttgtactgg
     4741 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     4801 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     4861 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcac
     4921 ccgggcgatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc
     4981 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5041 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5101 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5161 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5221 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5281 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5341 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     5401 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5461 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5521 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5581 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5641 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5701 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5761 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5821 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5881 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5941 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6001 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6061 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6121 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6181 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6241 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6301 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6361 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6421 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6481 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6541 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6601 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6661 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6721 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6781 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6841 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6901 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6961 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca
     7021 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     7081 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc
     7141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     7201 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7261 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa
     7321 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7381 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     7441 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     7501 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     7561 t
//
