LOCUS       sequence-335624-        3778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP green fluorescent protein CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3778)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3778)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: psfGFP
FEATURES             Location/Qualifiers
     source          1..3778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      494..503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      498..507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(537..556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(538..556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(563..580)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(563..579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(572..594)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      720..1175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(807..826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1017..1038
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1253..1357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1358..1426,1427..2218)
                     /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(1576..1595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2389..2977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2878..2897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3131..3148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3265..3286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3301..3306,3307..3324,3325..3331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3339..3355
                     /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     3344..3366
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3363..3379
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3363..3379
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3397..3415
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3397..3414
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gatgtacacg ttgtggctgt tgaagttgta ctccagcttg tggcccagga tgttgccatc
       61 ctccttgaaa tcgatgccct tcagctcgat gcggttcacc agggtatcgc cctcgaactt
      121 cacctcggcg cgggtcttgt aggtgccgtc atccttgaag ctgatggtgc gctcctgcac
      181 gtagccctcg ggcatggcgc tcttgaagaa atcgtgctgc ttcatgtgat cggggtagcg
      241 gctgaagcac tgcacgccgt aggtcagggt ggtcaccagg gtgggccagg gcacgggcag
      301 cttgccggtg gtgcagatga acttcagggt cagcttgccg ttggtggcgt cgccctcgcc
      361 ctcgccgcgc acgctgaact tgtggccgtt cacgtcgcca tccagctcca ccaggatggg
      421 caccacgccg gtgaacagct cctcgccctt ggacaccatt cgagaccact catcgcccaa
      481 tcgaattccc gcggccgcca tggcggccgg gagcatgcga cgtcgggccc aattcgccct
      541 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      601 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      661 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
      721 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      781 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      841 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
      901 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
      961 agcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1021 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1081 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1141 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     1201 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     1261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1321 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1381 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     1441 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     1501 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1561 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1621 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1681 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1741 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1801 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1861 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1921 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1981 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2041 tattgctgat aaatctggag ccggtgagcg tgggtcgcgc ggtatcattg cagcactggg
     2101 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2161 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2221 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2281 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2341 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3001 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3061 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3121 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3181 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3241 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     3301 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     3361 cacaggaaac agctatgacc atgattacgc caagctattt aggtgacact atagaatact
     3421 caagctatgc atccaacgcg ttgggagctc tcccatatgg tcgacctgca ggcggccgcg
     3481 aattcactag tgattgggcg atgagtggtc tcgaagctta cttgtacagc tcatccatgc
     3541 ccagggtgat gccggcggcg gtcacgaact ccagcagcac catgtgatcg cgcttctcgt
     3601 tggggtcctt ggacagcacg ctctgggtgc tcaggtagtg gttatcgggc agcagcactg
     3661 ggccgtcgcc gatgggggtg ttctgctggt agtgatcggc cagctgcacg gagccatcct
     3721 ccacattgtg gcggatcttg aagttggcct tgatgccgtt cttctgctta tcggcggt
//
