LOCUS       sequence-259128-        6843 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 6843)
  AUTHORS   Devroe E, Engelman A, Silver PA
  TITLE     Intracellular transport of human immunodeficiency virus type 1 
            integrase.
  JOURNAL   J Cell Sci. 2003 Nov 1. 116(Pt 21):4401-8.
  PUBMED    13130095
REFERENCE   2  (bases 1 to 6843)
  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: "J Cell Sci.
            2003 Nov 1. 116(Pt 21):4401-8."
COMMENT     Sequence Label: pPS2986
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..416
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        417..620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     571..591
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3683..3804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3720..3739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3774..3793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3811..4266)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3948..3969)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4160..4179
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4293..4397
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4399..4756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4399..4590
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4422..4442)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4607..4742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4791..5585
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4791..5585
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4845..4864)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5455..5474
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5773..5792)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5817..5864
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6193..6781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6682..6701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
       61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      601 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtactcagat ctcgagctca agcttcgaat taattcccac gctgccatgg cagacacctt
     1441 tctggagcac atgtgccgcc tggacatcga ctccgagcca accattgcca gaaacaccgg
     1501 catcatctgc accatcggcc cagcctcccg ctctgtggac aagctgaagg aaatgattaa
     1561 atctggaatg aatgttgccc gcctcaactt ctcgcacggc acccacgagt atcatgaggg
     1621 cacaattaag aacgtgcgag aggccacaga gagctttgcc tctgacccga tcacctacag
     1681 acctgtggct attgcactgg acaccaaggg acctgaaatc cgaactggac tcatcaaggg
     1741 aagtggcaca gcagaggtgg agctcaagaa gggcgcagct ctcaaagtga cgctggacaa
     1801 tgccttcatg gagaactgcg atgagaatgt gctgtgggtg gactacaaga acctcatcaa
     1861 agttatagat gtgggcagca aaatctatgt ggatgacggt ctcatttcct tgctggttaa
     1921 ggagaaaggc aaggactttg tcatgactga ggttgagaac ggtggcatgc ttggtagtaa
     1981 gaagggagtg aacctcccag gtgctgcggt cgacctgcct gcagtctcag agaaggacat
     2041 tcaggacctg aaatttggcg tggagcagaa tgtggacatg gtgttcgctt ccttcatccg
     2101 caaagctgct gatgtccatg ctgtcaggaa ggtgctaggg gaaaagggaa agcacatcaa
     2161 gattatcagc aagattgaga atcacgaggg tgtgcgcagg tttgatgaga tcatggaggc
     2221 cagcgatggc attatggtgg cccgtggtga cctgggtatt gagatccctg ctgaaaaagt
     2281 cttcctcgca cagaagatga tgattgggcg ctgcaacagg gctggcaaac ccatcatttg
     2341 tgccactcag atgttggaaa gcatgatcaa gaaacctcgc ccgacccgcg ctgagggcag
     2401 tgatgttgcc aatgcagttc tggatggagc agactgcatc atgctgtctg gggagaccgc
     2461 caagggagac tacccactgg aggctgtgcg catgcagcac gctattgctc gtgaggctga
     2521 ggccgcaatg ttccatcgtc agcagtttga agaaatctta cgccacagtg tacaccacag
     2581 ggagcctgct gatgccatgg cagcaggcgc ggtggaggcc tcctttaagt gcttagcagc
     2641 agctctgata gttatgaccg agtctggcag gtctgcacgc ctggtgtccc ggtaccgctt
     2701 cctggacggc atcgacaagg cccaggagga gcacgagaag taccactcca actggcgcgc
     2761 catggcctcc gacttcaacc tgccccccgt ggtggccaag gagatcgtgg cctcctgcga
     2821 caagtgccag ctgaagggcg aggccatgca cggccaggtg gactgctccc ccggcatctg
     2881 gcagctggac tgcacccacc tggagggcaa ggtgatcctg gtggccgtgc acgtggcctc
     2941 cggctacatc gaggccgagg tgatccccgc cgagaccggc caggagaccg cctacttcct
     3001 gctgaagctg gccggccgct ggcccgtgaa gaccgtgcac accgacaacg gctccaactt
     3061 cacctccacc accgtgaagg ccgcctgctg gtgggccggc atcaagcagg agttcggcat
     3121 cccctacaac ccccagtccc agggcgtgat cgagtccatg aacaaggagc tgaagaagat
     3181 catcggccaa gtccgcgacc aggccgagca cctgaagacc gccgtgcaga tggccgtgtt
     3241 catccacaac ttcaagcgca agggcggcat cggcggctac tccgccggcg agcgcatcgt
     3301 ggacatcatc gccaccgaca tccagaccaa ggagctgcag aagcagatca ccaagatcca
     3361 gaacttccgc gtgtactacc gcgactcccg cgaccccgtg tggaagggcc ccgccaagct
     3421 gctgtggaag ggcgagggcg ccgtggtgat ccaggacaac tccgacatca aggtggtgcc
     3481 ccgccgcaag gccaagatca tccgcgacta cggcaagcag atggccggcg acgactgcgt
     3541 ggcctcccgc caggacgagg actaatctag ataactgatc ataatcagcc ataccacatt
     3601 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3661 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3721 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3781 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     3841 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3901 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3961 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4021 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4081 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4141 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4201 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4261 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4321 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4381 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4561 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4621 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4681 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4741 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4801 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4861 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4921 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4981 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5041 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5101 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5161 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5221 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5281 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5341 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5401 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5461 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5521 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5581 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5641 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5701 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5761 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5821 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5881 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5941 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6001 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6061 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6121 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6301 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6841 gtt
//
