LOCUS       Exported                7032 bp DNA     circular SYN 11-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   hfytfiu
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7032)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 11, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(873..7032,1..872)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        410..650
                     /label=U6 Promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    703..820
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        869..1379
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             1401..2000
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             2128..2361
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2433..2554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2594..2729
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2750..2768)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2778..2794)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2936..3391
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3417..3521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3522..4382
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4553..5141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5429..5450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5465..5495
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5503..5519
                     /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     5527..5543
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5564..5582
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5608..5836
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5837..6017
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6064..6189
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6682..6915
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
       61 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
      121 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
      181 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
      241 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
      301 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
      361 tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg agggcctatt
      421 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      481 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      541 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      601 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
      661 tgaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
      721 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      781 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
      841 cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc caaggcagcc
      901 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
      961 accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta
     1021 cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc
     1081 ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag
     1141 acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg
     1201 gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt
     1261 gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc
     1321 cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg
     1381 gggatccacc ggagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     1441 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     1501 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     1561 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     1621 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     1681 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     1741 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     1801 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     1861 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     1921 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     1981 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     2041 cacgacccca tgcatcggta cctttaagac caatgactta caaggcagct gtagatctta
     2101 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     2161 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     2221 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     2281 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     2341 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     2401 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     2461 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2521 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     2581 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2641 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2701 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     2761 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2821 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2881 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     2941 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3001 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3061 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3121 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3181 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3241 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3301 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3361 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     3421 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3481 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3541 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3601 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3661 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3721 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3781 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3841 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3901 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3961 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4021 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4081 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4141 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4201 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4261 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4321 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4381 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4441 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4501 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4561 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4621 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4681 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4741 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4801 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4861 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4921 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4981 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5041 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5101 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5161 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5221 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5281 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5341 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5401 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     5461 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     5521 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     5581 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
     5641 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
     5701 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
     5761 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
     5821 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5881 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5941 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6001 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     6061 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     6121 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     6181 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     6241 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     6301 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     6361 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     6421 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     6481 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     6541 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     6601 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     6661 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     6721 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     6781 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     6841 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     6901 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     6961 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     7021 gatggagtgg ga
//
