Geographic Information System has extended to higher assessment of water resources. GIS linking with hydrological model becomes a trend in water resource assessment modeling. One of the most popular models is SWAT2000 which have effectiveness in multi-purpose processes for predicting the impact of land management practices on water, sediments and chemicals yields in large complex watershed with varying soils, land uses, and management conditions over long period of time. In this study, SWAT2000 model was applied to Gap stream watershed in Daejeon city where TMDL (Total Maximum Daily Load) Regulation would be implanted. The Gap Stream watershed was partitioned into 8 subbasins, however, only 3 out of 8 subbaisns were observed for having practical gauged data on the basis of streamflow from the year of 2002 to 2005. Gauged streamflow data of Indong, Boksu and Hoeduck stations were used for calibration and validation of the SWAT Streamflow simulation. Estimation Efficiency Analysis (COE), Regression Analysis ($R^{2}$), Relative Error (R.E.) were used for comparing observed streamflow data of the 3 subbasins on the daily and monthly basis with estimated streamflow data in order to fix optimized parameters for the best fitted results. COE value for the daily and monthly streamflow was ranged from 0.45 to 0.96. $R^{2}$ values for daily and monthly streamflow ranged from 0.51 to 0.97. R.E. values for total streamflow volume ranged from 3 % to 22.5 %. The accuracy of the model results shows that the SWAT2000 model can be applicable to Korean watersheds like the Gap Stream watershed that needs to be partitioned into a number of subbasins for TMDL regulation.