LOCUS       sequence-402893-        7640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes GFP alone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7640)
  AUTHORS   Roman-Fernandez A, Mansour MA, Kugeratski FG, Anand J, Sandilands E,
            Galbraith L, Rakovic K, Freckmann EC, Cumming EM, Park J, Nikolatou 
            K, Lilla S, Shaw R, Strachan D, Mason S, Patel R, McGarry L, Katoch 
            A, Campbell KJ, Nixon C, Miller CJ, Leung HY, Le Quesne J, Norman 
            JC, Zanivan S, Blyth K, Bryant DM
  TITLE     Spatial regulation of the glycocalyx component podocalyxin is a 
            switch for prometastatic function.
  JOURNAL   Sci Adv. 2023 Feb 3;9(5):eabq1858. doi: 10.1126/sciadv.abq1858. Epub
            2023 Feb 3.
  PUBMED    36735782
REFERENCE   2  (bases 1 to 7640)
  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: "Sci Adv."; 
            date: "2023-02-3"; volume: "9(5):eabq1858. doi"; pages: " 
            10.1126/sciadv.abq1858. Epub 2023 Feb 3"
COMMENT     Sequence Label: pQCXIZ GFP C1
FEATURES             Location/Qualifiers
     source          1..7640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1604..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2108..2132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2257..2266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2263..2265,2266..2268,2269..2979)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2263..2265,2266..2268,2269..2979)
                     /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(2308..2329)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2569..2588)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2916..2937
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2980..3045
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3276..3738
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3332..3349)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3559..3578
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3752..4126
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             4346..4771
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4874..4896)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5032..5052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5037..5366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5217..5352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5388..5407
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5488..5505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5659..6247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5739..5758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6418..7209,7210..7278))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7041..7060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7279..7383)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7451..7469
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7569..7640,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgggggatcg atcctctaga gtccgttaca taacttacgg
     1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1921 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2101 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2161 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
     2221 cggacctgca ggaattgatc cgcggccgca ccggtcgcca ccatggtgag caagggcgag
     2281 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2341 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2401 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2461 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2521 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2581 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2641 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2701 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2761 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2821 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2881 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2941 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct
     3001 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa
     3061 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3121 cacctccccc tgaacctgaa acataaaatg aatgcaattc csccccccyc yccctccccc
     3181 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     3241 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     3301 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     3361 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     3421 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     3481 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     3541 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     3601 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     3661 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     3721 tgaaaaacac gatgataagc ttgccacaac catggccaag ttgaccagtg ccgttccggt
     3781 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     3841 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     3901 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     3961 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     4021 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     4081 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg acccggacgg
     4141 gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc
     4201 cgttttccgc ctgaggtcac tgcgtggatg gagcatcagt ggtccaggct ctagttttga
     4261 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     4321 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4381 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4441 gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag atgtttccag
     4501 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     4561 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     4621 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     4681 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     4741 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     4801 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg
     4861 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4921 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4981 gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt
     5041 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5101 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5161 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5221 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5281 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5341 tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg catcagagca
     5401 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5461 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6061 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6541 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6721 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7021 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7381 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7441 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag cttggccatt
     7501 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     7561 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7621 tcatagccca tatatggagt
//
