LOCUS       pRK5-mEGFP-FOXF1        6524 bp DNA     circular SYN 16-NOV-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Nov 16, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      933..941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             939..1648
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1:  939 ..  941 / #05fd14
                        2:  942 ..  944 / #05fd14 / 1a
                        3:  945 .. 1648 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDE"
     CDS             1659..2798
                     /codon_start=1
                     /label=FOXF1_wt
                     /note="color: #993366"
                     /translation="MSSAPEKQQPPHGGGGGGGGGGGAAMDPASSGPSKAKKTNAGIRR
                     PEKPPYSYIALIVMAIQSSPTKRLTLSEIYQFLQSRFPFFRGSYQGWKNSVRHNLSLNE
                     CFIKLPKGLGRPGKGHYWTIDPASEFMFEEGSFRRRPRGFRRKCQALKPMYSMMNGLGF
                     NHLPDTYGFQGSAGGLSCPPNSLALEGGLGMMNGHLPGNVDGMALPSHSVPHLPSNGGH
                     SYMGGCGGAAAGEYPHHDSSVPASPLLPTGAGGVMEPHAVYSGSAAAWPPSASAALNSG
                     ASYIKQQPLSPCNPAANPLSGSLSTHSLEQPYLHQNSHNAPAELQGIPRYHSQSPSMCD
                     RKEFVFSFNAMASSSMHSAGGGSYYHQQVTYQDIKPCVM"
     polyA_signal    2832..2966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     promoter        3039..3396
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3247..3382
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     complement(3416..3432)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      3645..4100
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4382..4486
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4487..5347
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4487 .. 4555 / #ccffcc / signal sequence
                        2: 4556 .. 5347 / #ccffcc
                     Cleavage site after base 4555"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5518..6106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        6430..6460
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 6430 .. 6435 / #ffffff / -35
                        2: 6436 .. 6453 / #ffffff
                        3: 6454 .. 6460 / #ffffff / -10"
     protein_bind    6468..6484
                     /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)."
                     /note="color: #31849b"
     primer_bind     6492..6508
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgtaccg gtgccaccat ggtgagcaag ggcgaggagc
      961 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1021 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     1081 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1141 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1201 ccatgcccga aggctacgtc caggagcgca ccatctcctt caaggacgac ggcacctaca
     1261 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1321 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca
     1381 gccacaacgt ctatatcacg gccgacaagc agaagaacgg catcaaggcg aacttcaaga
     1441 tccgccacaa cgtcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1501 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     1561 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1621 ccgggatcac tctcggcatg gacgagctgt acacggcgat gagtagcgcc cccgagaaac
     1681 aacaaccacc gcatggagga ggaggaggcg gcggtggtgg tggtggggct gcaatggatc
     1741 cggcatccag tggaccctca aaagcgaaga aaacaaatgc aggaataagg cggcctgaaa
     1801 agccaccgta ctcttatatt gcccttattg tgatggctat ccaaagctct ccaactaaaa
     1861 ggctgacctt gtccgaaatt tatcaatttc ttcaatctag atttcctttc tttcgaggga
     1921 gctatcaagg gtggaagaat agtgtccggc ataatttgag cctgaatgaa tgttttatta
     1981 aattaccaaa aggtctcggc cgacctggga aagggcatta ttggacaata gatcctgcat
     2041 cagaatttat gtttgaagaa gggtcattca ggcgaagacc acggggattt cggagaaagt
     2101 gtcaagcact gaaaccgatg tattccatga tgaatggcct gggatttaat catcttcccg
     2161 atacttatgg ttttcaaggg agcgcaggtg ggcttagctg tccaccaaat agtctcgcct
     2221 tggaaggcgg gctcgggatg atgaatggac atcttcctgg aaatgtagat ggaatggctc
     2281 ttccatctca ttctgtccca catcttccca gtaatggtgg tcattcttat atgggcggtt
     2341 gtggcggagc cgccgcaggg gaatatcccc atcatgatag ttctgtcccg gcttctccac
     2401 tccttccaac aggagcaggc ggcgtaatgg aaccccatgc ggtttattcc gggtctgctg
     2461 ctgcatggcc ccctagcgcc tctgccgccc tgaattccgg agcaagctac ataaaacaac
     2521 aaccactctc tccatgcaat ccggcagcta atcctttgag cgggtccctg tcaacccata
     2581 gcttagaaca accctacttg catcaaaatt cacataatgc tcccgctgaa ctccagggga
     2641 taccacgcta ccatagccaa tccccttcta tgtgcgatcg taaagaattc gtgttttcct
     2701 ttaatgcgat ggcaagctcc tctatgcatt ctgcgggcgg tgggagctat tatcatcaac
     2761 aagtgactta tcaggatatt aaaccctgtg ttatgtaaag tcgacctgca gaagcttggc
     2821 cgccatggcc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2881 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2941 tcaatgtatc ttatcatgtc tggatcgatc gggaattaat tcggcgcagc accatggcct
     3001 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     3061 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3121 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3241 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3301 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3361 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg
     3421 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3481 cagcacatcc ccccttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3541 cccaacagtt gcgtagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     3601 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     3661 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3721 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3781 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3841 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3901 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3961 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4021 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4081 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     4141 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4201 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4261 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4321 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4381 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4441 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4501 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4561 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4621 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4681 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg atgacgccgg
     4741 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4801 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4861 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4921 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4981 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgccag cagcaatggc
     5041 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5101 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5161 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5221 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5281 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5341 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5401 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5461 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5521 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5581 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5641 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5701 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5761 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5821 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5881 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     5941 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6001 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6061 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6241 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6301 caaaccgcct ctccccgcgc gttggccgat tcattaatcc agctggcacg acaggtttcc
     6361 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttacctca ctcattaggc
     6421 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6481 acaatttcac acaggaaaca gctatgacca tgattacgaa ttaa
//
