LOCUS       sequence-11363-e        7129 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 7129)
  AUTHORS   Brugmann SA, Goodnough LH, Gregorieff A, Leucht P, ten Berge D, 
            Fuerer C, Clevers H, Nusse R, Helms JA
  TITLE     Wnt signaling mediates regional specification in the vertebrate 
            face.
  JOURNAL   Development. 2007 Sep . 134(18):3283-95.
  PUBMED    17699607
REFERENCE   2  (bases 1 to 7129)
  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: 
            "Development. 2007 Sep . 134(18):3283-95."
COMMENT     Sequence Label: 7TG
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        201..232
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      288..297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(294..296,297..299,300..1013)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(294..296,297..299,300..1010)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(339..360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(600..619)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     947..968
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1038..1626
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1091..1111)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1509..1520)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1713..1946
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2018..2139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2055..2074)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2109..2128
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2158..2293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2325..2344)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2326..2344)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2354..2371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2354..2370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2515..2970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2602..2621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2812..2833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2999..3103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3104..3172,3173..3964)
                     /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(3322..3341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4135..4723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4624..4643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4877..4894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5011..5032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5047..5052,5053..5070,5071..5077)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5085..5101
                     /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     5090..5112
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5109..5125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5109..5125
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5144..5164
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5146..5164
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             5420..5600
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5644..5769
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6262..6495
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6680..6724
                     /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"
     misc_feature    6991..7108
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctagcccggg ctcgagcagc tgaagcttgc atgcctgcag ggtaccgagc tcttacgcga
       61 gatcaaaggg ggtaagatca aagggggtaa gatcaaaggg gcgcgagatc aaagggggta
      121 agatcaaagg gggtaagatc aaagggggta agatcaaagg ggcgcgcccg cgtgctagcc
      181 cgggctcgag atctagactc tagagggtat ataatggaag ctcgaattcc agcttggcat
      241 tccggtactg ttggtaaaaa gcttggcatt ccggtactgt tggtaaagcc accatggtga
      301 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      361 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      421 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      481 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      541 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      601 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      661 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      721 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
      781 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
      841 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
      901 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
      961 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     1021 gcgactctag agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     1081 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     1141 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     1201 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     1261 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     1321 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     1381 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     1441 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     1501 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     1561 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     1621 ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     1681 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     1741 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     1801 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     1861 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     1921 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     1981 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     2041 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2101 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     2161 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2221 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2281 ttttttggag gcctaggctt ttgcgtcgag acgtacccaa ttcgccctat agtgagtcgt
     2341 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2401 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc
     2521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2581 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2641 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2701 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2821 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2881 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2941 attttaacaa aatattaacg tttacaattt cccaggtggc acttttcggg gaaatgtgcg
     3001 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3061 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3121 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3181 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3241 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3301 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3361 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3421 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3481 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3541 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3601 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3661 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3721 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3781 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3841 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3901 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3961 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4021 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4081 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4141 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4201 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4261 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4321 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4381 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4441 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4501 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4561 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4621 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4681 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4741 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4801 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4861 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4921 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     4981 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5041 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5101 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     5161 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     5221 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     5281 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     5341 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     5401 ctagctcgat acaataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     5461 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5521 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5581 agtgtggaaa atctctagca gtggcgcccg aacagggacc tgaaagcgaa agggaaacca
     5641 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     5701 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     5761 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     5821 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     5881 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     5941 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     6001 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     6061 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     6121 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     6181 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     6241 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     6301 cactatgggc gcagcctcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     6361 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     6421 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     6481 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     6541 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     6601 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     6661 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     6721 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     6781 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     6841 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     6901 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     6961 agtgaacgga tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta
     7021 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     7081 aaaacaaatt acaaaaattc aaaattttat cgatcacgag actagcctc
//
