LOCUS       Exported                7547 bp ds-DNA     circular SYN 01-NOV-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7547)
  AUTHORS   CSHL
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 4, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7547
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..413
                     /label=ApEinfo_fwdcolor=#bc8dbf_ApEinfo_revcolor=#bc8d
                     /label=ApEinfo_fwdcolor=#bc8dbf_ApEinfo_revcolor=#bc8d...
                     /note="label=""5'LTR"" - note=""ApEinfo_revcolor=#bc8dbf""
                     - note=""ApEinfo_fwdcolor=#bc8dbf"" - 1..413"
     LTR             232..412
                     /label=HIV-1 5 LTR
     misc_feature    464..601
                     /label=psi
                     /note="label=""psi"" - 464..601"
     misc_feature    523..567
                     /label=HIV-1 psi pack
     misc_feature    568..932
                     /label=gag
                     /note="label=""gag"" - 568..932"
     misc_feature    1077..1310
                     /label=RRE
     misc_feature    1078..1319
                     /label=RRE
                     /note="label=""RRE"" - 1078..1319"
     misc_feature    1694..1713
                     /label=pre-hU6_SEQ
     promoter        1717..1941
                     /label=hU6
                     /note="label=""hU6"" - 1717..1965"
     misc_feature    1717..1941
                     /label=hU6 promoter
     misc_feature    1717..1739
                     /label=SANGER SEQ primer
     misc_feature    1884..1924
                     /label=pool screening F_misc_feature
                     /note="note=""pool screening F"" - 1884..1924"
     misc_feature    1917..1941
                     /label=cloning_fwd_primer
     misc_feature    2420..2502
                     /label=sgRNA backbone_misc_feature
                     /note="note=""sgRNA backbone"" - 3847..3949"
     misc_feature    2420..2502
                     /label=sg_scaffold_LRG
     misc_feature    2420..2449
                     /label=cloning_rev_primer
     misc_feature    2544..2661
                     /label=cPPT
                     /note="label=""cPPT"" - 3991..4108"
     misc_feature    2573..2597
                     /label=pool screening R (2nd round)_misc_feature
                     /note="note=""pool screening R (2nd round)"" - 4020..4044"
     misc_feature    2583..2626
                     /label=pool screening R_misc_feature
                     /note="note=""pool screening R"" - 4030..4073"
     promoter        2705..2955
                     /label=EFS
                     /note="label=""EFS"" - 4152..4402"
     misc_feature    2705..2955
                     /label=EFS1 promoter
     misc_feature    2730..2941
                     /label=EFS_core_promoter
     misc_feature    complement(2805..2810)
                     /label=Kozak-site
     misc_feature    2961..2966
                     /label=Kozak-site
     misc_feature    2966..3389
                     /label=GFP_misc_feature [*]
                     /note="note=""GFP"" - 4441..5160"
     misc_feature    2967..3386
                     /label=BlastR
     misc_binding    3404..3992
                     /label=WPRE
                     /note="bound_moiety="""" - label=""WPRE"" - 5175..5763"
     misc_feature    3404..3992
                     /label=WPRE
     misc_feature    3656..3661
                     /label=Kozak-site
     misc_feature    3833..3838
                     /label=Kozak-site
     CDS             complement(3875..3886)
                     /label=FactorXa site
     LTR             4063..4298
                     /label=3' LTR SIN
                     /note="label=""3' LTR SIN"" - 5834..6069"
     misc_feature    4064..4116
                     /label=delta U3
     LTR             4117..4297
                     /label=HIV-1 5 LTR
     polyA_signal    4375..4505
                     /label=SV40 polyadenylation signal
                     /note="label=""SV40 polyadenylation signal"" - 6146..6276"
     misc_feature    4536..4613
                     /label=SV40 ORI
     rep_origin      4585..4660
                     /direction=RIGHT
                     /label=SV40 origin of replication
                     /note="label=""SV40 origin of replication"" - 6356..6431"
     primer_bind     complement(4691..4710)
                     /label=T7
     primer_bind     complement(4720..4737)
                     /label=M13-fwd
     CDS             4808..4876
                     /label=LacZ alpha
     rep_origin      4878..5333
                     /direction=RIGHT
                     /label=F1 ori
                     /note="label=""F1 ori"" - 6649..7104"
     rep_origin      complement(4883..5323)
                     /direction=LEFT
                     /label=F1 ori
     misc_feature    complement(5335..5340)
                     /label=Kozak-site
     promoter        5394..5422
                     /label=Amp prom
     CDS             5464..6324
                     /label=AmpR
                     /note="label=""AmpR"" - 7235..8095"
     CDS             5662..6321
                     /label=AmpR
     rep_origin      6472..7139
                     /direction=RIGHT
                     /label=pUC ori
                     /note="label=""pUC ori"" - 8243..8910"
     rep_origin      6473..7101
                     /direction=RIGHT
                     /label=ColE1 origin
     misc_feature    6665..6670
                     /label=Kozak-site
     misc_feature    7022..7027
                     /label=Kozak-site
     promoter        7407..7436
                     /label=lac
     misc_binding    7441..7463
                     /label=LacO
     primer_bind     7469..7489
                     /label=M13-rev
     primer_bind     7507..7526
                     /label=T3
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgtctt agagaaactt
     1981 ggagagtttc agagctatgc acaagagtgc atagcaagtt gaaataaggc tagtccgtta
     2041 tcaactttgc tggaaacagc aaagtggcac cgagtcggtg ctttttttgc gtgaccgagc
     2101 ttgtctgccc gcgcagtcca ctagacagac gcgcagaggt cgggcgggcc tagggccaat
     2161 ttcccatggt tcccttcatt tgcatatatg atgtaataag gttatggaga ctattaaact
     2221 tagccctaat caaactatat gatgataatg tgtgtggtac aaaaggtcat aacttattat
     2281 atactttgaa acttaaaaaa gggttacagt ttagtcacca taactgtaaa attttttcta
     2341 ttcttagctt tatatagttc ttgagaggcc atgtttatcg gatgtatatc agcataaggg
     2401 tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact
     2461 ttgctggaaa cagcaaagtg gcaccgagtc ggtgcttttt ttgaattcta gatcttgaga
     2521 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     2581 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     2641 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc
     2701 cggctcgagt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     2761 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     2821 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     2881 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     2941 ggtgtcgtga cgcgggatcc gccaccatga agaccttcaa catctctcag caggaccttg
     3001 agctggtgga agttgccact gagaaaatca ccatgctcta tgaggacaac aagcaccatg
     3061 ttggtgctgc catcaggacc aagacaggag aaatcatctc tgctgtccac attgaagcct
     3121 acattggcag ggtcactgtg tgtgcagagg ccattgccat tgggtctgca gtctccaatg
     3181 ggcagaagga ctttgacacc attgtggctg tgaggcaccc ctacagtgat gaggtggaca
     3241 ggtccatcag agtggtgtcc ccctgtggca tgtgcaggga gctcatctca gactatgccc
     3301 ctgattgctt tgttctgatt gagatgaatg gcaagctggt caagacaacc attgaggagc
     3361 tgattccact gaaatacacc agaaactaac gcgttaagtc gacaatcaac ctctggatta
     3421 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3481 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3541 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3601 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3661 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3721 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3781 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     3841 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3901 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3961 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     4021 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     4081 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     4141 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     4201 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     4261 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     4321 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     4381 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4441 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4501 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4561 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4621 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     4681 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     4741 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4801 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4861 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4921 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4981 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5041 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5101 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5161 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5221 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5281 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5341 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5401 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5461 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5521 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5581 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5641 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5701 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5761 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5821 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5881 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5941 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6001 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6061 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6121 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6181 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6241 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6301 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6361 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6421 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6481 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6541 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6601 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6661 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6721 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6781 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6841 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6901 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6961 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7021 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7081 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7141 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7201 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7261 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7321 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7381 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7441 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7501 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagc
//
