LOCUS       sequence-460241-        7776 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 7776)
  AUTHORS   Scott KL, Nogueira C, Heffernan TP, van Doorn R, Dhakal S, Hanna JA,
            Min C, Jaskelioff M, Xiao Y, Wu CJ, Cameron LA, Perry SR, Zeid R, 
            Feinberg T, Kim M, Vande Woude G, Granter SR, Bosenberg M, Chu GC, 
            DePinho RA, Rimm DL, Chin L
  TITLE     Proinvasion metastasis drivers in early-stage melanoma are 
            oncogenes.
  JOURNAL   Cancer Cell. 2011 Jul 12;20(1):92-103.
  PUBMED    21741599
REFERENCE   2  (bases 1 to 7776)
  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: "Cancer 
            Cell."; date: "2011-07-12"; volume: "20(1)"; pages: "92-103"
COMMENT     Sequence Label: pLenti6/V5-DEST-MTHFD2
FEATURES             Location/Qualifiers
     source          1..7776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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"
     enhancer        1814..2117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2118..2321
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2272..2292
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2318..2342
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2438..2462
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3207..3231)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3284..3325
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3362..3382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3367..3696
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3547..3682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3744..3791
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3810..4208
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4295..4528
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4600..4721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4637..4656)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4691..4710
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4761..4896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4916..4935)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4917..4935)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4945..4962)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4945..4961)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4954..4976)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5103..5558
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5190..5209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5400..5421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5584..5688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5689..5757,5758..6549)
                     /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(5907..5926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6720..7308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7209..7228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7462..7479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7596..7617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7632..7637,7638..7655,7656..7662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7670..7686
                     /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     7675..7697
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7694..7710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7694..7710
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7729..7749
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7731..7749
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgataag
     1801 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     1861 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     1921 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1981 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2041 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2101 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2161 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2221 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2281 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     2341 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagagga
     2401 tccactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca aaaaagttgg
     2461 catgaaacca gcttcaattt cagaggaaga attgttgaat ttaatcaata aactgaataa
     2521 tgatgataat gtagatggcc tccttgttca gttgcctctt ccagagcata ttgatgagag
     2581 aaggatctgc aatgctgttt ctccagacaa ggatgttgat ggctttcatg taattaatgt
     2641 aggacgaatg tgtttggatc agtattccat gttaccggct actccatggg gtgtgtggga
     2701 aataatcaag cgaactggca ttccaaccct agggaagaat gtggttgtgg ctggaaggtc
     2761 aaaaaacgtt ggaatgccca ttgcaatgtt actgcacaca gatggggcgc atgaacgtcc
     2821 cggaggtgat gccactgtta caatatctca tcgatatact cccaaagagc agttgaagaa
     2881 acatacaatt cttgcagata ttgtaatatc tgctgcaggt attccaaatc tgatcacagc
     2941 agatatgatc aaggaaggag cagcagtcat tgatgtggga ataaatagag ttcacgatcc
     3001 tgtaactgcc aaacccaagt tggttggaga tgtggatttt gaaggagtca gacaaaaagc
     3061 tgggtatatc actccagttc ctggaggtgt tggccccatg acagtggcaa tgctaatgaa
     3121 gaataccatt attgctgcaa aaaaggtgct gaggcttgaa gagcgagaag tgctgaagtc
     3181 taaagagctt ggggtagcca ctaattaccc aactttcttg tacaaagtgg ttgatatcca
     3241 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3301 ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttgga attaattctg
     3361 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3421 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3541 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3601 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3661 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3721 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     3781 tataatacga caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca
     3841 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     3901 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     3961 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     4021 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     4081 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     4141 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     4201 agggctaagc acaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta
     4261 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     4321 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     4381 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     4441 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     4501 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     4561 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     4621 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4681 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     4741 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4801 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4861 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     4921 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4981 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5041 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5101 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5161 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5221 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5281 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5341 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5401 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5461 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5521 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     5581 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5641 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5701 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5761 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5821 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5881 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5941 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6001 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6061 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6121 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6181 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6241 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6301 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6361 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6421 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6481 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6541 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6601 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6661 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6721 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6781 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6841 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6901 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6961 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7021 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7081 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7141 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7201 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7261 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7321 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7381 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7441 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7501 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7561 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7621 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7681 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     7741 cactaaaggg aacaaaagct ggagctgcaa gcttaa
//
