LOCUS       sequence-263313-        7085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7085)
  AUTHORS   Orimo A, Gupta PB, Sgroi DC, Arenzana-Seisdedos F, Delaunay T, Naeem
            R, Carey VJ, Richardson AL, Weinberg RA
  TITLE     Stromal fibroblasts present in invasive human breast carcinomas 
            promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 
            secretion.
  JOURNAL   Cell. 2005 May 6. 121(3):335-48.
  PUBMED    15882617
REFERENCE   2  (bases 1 to 7085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2005 
            May 6. 121(3):335-48."
COMMENT     Sequence Label: pLKO.1 puro CXCR4 siRNA-1
FEATURES             Location/Qualifiers
     source          1..7085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        1606..1854
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1606..1626
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1777..1796
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    1952..2069
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2118..2628
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2547..2565
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2650..3249
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2650..2669)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3146..3166
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3377..3610
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3682..3803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3719..3738)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3773..3792
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3843..3978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3998..4017)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3999..4017)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4027..4044)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4027..4043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4036..4058)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4185..4640
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4272..4291)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4482..4503
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4666..4770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4771..4839,4840..5631)
                     /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"
     primer_bind     complement(4989..5008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5802..6390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6291..6310
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6544..6561
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6678..6699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6714..6719,6720..6737,6738..6744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6752..6768
                     /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     6757..6779
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6776..6792
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6776..6792
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6811..6831
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6813..6831
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6859..7085
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg cctatttccc
     1621 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     1681 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     1741 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     1801 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggttgg
     1861 aggggatcag tatatacact cgagtgtata tactgatccc ctccattttt gaattctcga
     1921 cctcgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     1981 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     2041 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
     2101 tttggccgcg gctcgagggg gttggggttg cgccttttcc aaggcagccc tgggtttgcg
     2161 cagggacgcg gctgctctgg gcgtggttcc gggaaacgca gcggcgccga ccctgggtct
     2221 cgcacattct tcacgtccgt tcgcagcgtc acccggatct tcgccgctac ccttgtgggc
     2281 cccccggcga cgcttcctgc tccgccccta agtcgggaag gttccttgcg gttcgcggcg
     2341 tgccggacgt gacaaacgga agccgcacgt ctcactagta ccctcgcaga cggacagcgc
     2401 cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca
     2461 gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg tggggcggta
     2521 gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg
     2581 tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg ggatccaccg
     2641 gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     2701 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     2761 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     2821 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     2881 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     2941 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     3001 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     3061 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     3121 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     3181 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     3241 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
     3301 gcatcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     3361 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     3421 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     3481 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     3541 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     3601 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     3661 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     3721 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3781 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     3841 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3901 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3961 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     4021 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     4081 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     4141 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     4201 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4261 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4321 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4381 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4441 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4501 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4561 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4621 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4981 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5281 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5941 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6421 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6481 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6541 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6601 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6661 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6721 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6781 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     6841 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     6901 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     6961 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     7021 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     7081 taaac
//
